Structured Review

Ribobio lncrna snhg7
<t>LncRNA-</t> <t>SNHG7</t> promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
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Images

1) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

2) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

3) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

4) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

5) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

6) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

7) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

8) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

9) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

10) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

11) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

12) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

13) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

14) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

15) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

16) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

17) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

18) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

19) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

20) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

21) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

22) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

23) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

24) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

25) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

26) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

27) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

28) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

29) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

30) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

31) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

32) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

33) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

34) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

35) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

36) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

37) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

38) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

39) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

40) Product Images from "c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer"

Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

Journal: Journal of Breast Cancer

doi: 10.4048/jbc.2019.22.e54

LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
Figure Legend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

Techniques Used: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p
Figure Legend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

Techniques Used: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p
Figure Legend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p
Figure Legend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

Techniques Used: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p
Figure Legend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

Techniques Used: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

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    Ribobio lncrna snhg7
    <t>LncRNA-</t> <t>SNHG7</t> promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p
    Lncrna Snhg7, supplied by Ribobio, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

    Journal: Journal of Breast Cancer

    Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

    doi: 10.4048/jbc.2019.22.e54

    Figure Lengend Snippet: LncRNA- SNHG7 promotes proliferation of breast cancer cells. (A, B) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then cells were subjected to cell proliferation analysis. (C) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus and then subjected to cell viability assays. (D, E) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus. LncRNA- SNHG7 level was assessed by qRT-PCR, and then the cells were subjected to cell proliferation analysis. (F) MCF-7 cells were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus, and then subjected to cell viability assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; sh = short hairpin. * p

    Article Snippet: Increased level of lncRNA-SNHG7 was detected in breast cancer tissue and cell lines To investigate the potential role of lncRNA-SNHG7 in breast cancer, we monitored the level of lncRNA-SNHG7 in breast cancer tissues.

    Techniques: Transduction, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

    LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

    Journal: Journal of Breast Cancer

    Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

    doi: 10.4048/jbc.2019.22.e54

    Figure Lengend Snippet: LncRNA- SNHG7 is a direct transcriptional target of c-Myc. (A, B) MCF-7 and MDA-MB-231 cells were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus. Western blot analysis of c-Myc and β-actin. LncRNA- SNHG7 level was assessed by quantitative RT-PCR. (C) Schematic illustration of the consensus c-Myc binding sites in lncRNA- SNHG7 gene promoter. The wild-type and mutant binding sites are shown in the open boxes. The corresponding pGL3-based luciferase reporter constructs generated are shown. (D, E) MCF-7 were transduced with sh-ctrl or sh-c-Myc lentivirus, pCDH or pCDH-c-Myc lentivirus followed by transfection with the indicated pGL3-based luciferase reporter constructs. The reporter activity was measured 24 hours post-transfection, and plotted after normalizing with the Renilla luciferase activity. Western blot analysis of c-Myc and β-actin. (F) MCF-7 cells lysates were analyzed by ChIP assay using anti-c-Myc or IgG rabbit antibody. The ChIP products were amplified by semi-quantitative RT-PCR. (G) Correlation analyses conducted between c-Myc and lncRNA- SNHG7 in breast cancers (n = 50 biologically independent samples). Pearson correlation coefficients ( r ) and p -values. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; RT-PCR = real-time polymerase chain reaction; NS = not significant; GAPDH = glyceraldehyde 3-phosphate dehydrogenase; IgG = immunoglobulin G. * p

    Article Snippet: Increased level of lncRNA-SNHG7 was detected in breast cancer tissue and cell lines To investigate the potential role of lncRNA-SNHG7 in breast cancer, we monitored the level of lncRNA-SNHG7 in breast cancer tissues.

    Techniques: Multiple Displacement Amplification, Transduction, Western Blot, Quantitative RT-PCR, Binding Assay, Mutagenesis, Luciferase, Construct, Generated, Transfection, Activity Assay, Chromatin Immunoprecipitation, Amplification, Standard Deviation, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

    Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

    Journal: Journal of Breast Cancer

    Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

    doi: 10.4048/jbc.2019.22.e54

    Figure Lengend Snippet: Elevated expression of lncRNA- SNHG7 in breast cancer. (A) qRT-PCR for the abundance of lncRNA- SNHG7 in 30 pairs of breast cancer samples. (B) The correlation between overall survival and lncRNA- SNHG7 level in breast cancer obtained by TANRIC. (C) qRT-PCR for the abundance of lncRNA- SNHG7 in normal breast cells and various breast cancer cell lines. (D) Western-blot analysis for PARP and β-actin in the nuclear and cytoplasmic fractions, respectively, in MCF-7 cells. Data shown represent 3 independent experiments. (E) Nuclear and cytoplasmic fractions of MCF-7 cells were subjected to qRT-PCR analysis. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; qRT-PCR = quantitative real-time polymerase chain reaction; PARP = poly (ADP-ribose) polymerase. * p

    Article Snippet: Increased level of lncRNA-SNHG7 was detected in breast cancer tissue and cell lines To investigate the potential role of lncRNA-SNHG7 in breast cancer, we monitored the level of lncRNA-SNHG7 in breast cancer tissues.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction

    Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

    Journal: Journal of Breast Cancer

    Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

    doi: 10.4048/jbc.2019.22.e54

    Figure Lengend Snippet: Knocking down lncRNA- SNHG7 inhibits glycolysis in breast cancer cells. (A) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Acidification of the culture medium was evaluated by visually inspecting the color of the medium. (B) MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA were cultured for 24 hours. Levels of lactate in the culture medium were then measured and normalized to cell number. (C, D) ECAR was measured in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA by Seahorse XF assays. (E, F) Western blotting and qRT-PCR analysis of glycolysis enzymes in MCF-7 cells expressing either control shRNA or lncRNA- SNHG7 shRNA. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction. * p

    Article Snippet: Increased level of lncRNA-SNHG7 was detected in breast cancer tissue and cell lines To investigate the potential role of lncRNA-SNHG7 in breast cancer, we monitored the level of lncRNA-SNHG7 in breast cancer tissues.

    Techniques: Expressing, shRNA, Cell Culture, Western Blot, Quantitative RT-PCR, Standard Deviation, Real-time Polymerase Chain Reaction

    LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

    Journal: Journal of Breast Cancer

    Article Title: c-Myc-Induced Long Non-Coding RNA Small Nucleolar RNA Host Gene 7 Regulates Glycolysis in Breast Cancer

    doi: 10.4048/jbc.2019.22.e54

    Figure Lengend Snippet: LncRNA- SNHG7 acts as a target of miR-34a-5p to increase LDHA level. (A) Illustration of the base pairing between miR-34a-5p and lncRNA- SNHG7 . The base pairing between miR-34a-5p and LDHA 3′UTR is also shown. (B) Lysates from MCF-7 cells were incubated with in vitro -synthesized biotin-labeled sense or antisense DNA probes against lncRNA- SNHG7 for biotin pull-down assay, followed by qRT-PCR analysis to examine miR-34a-5p and lncRNA- SNHG7 level. (C) MCF-7 were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. Western blot analysis of LDHA and β-actin. (D) MCF-7 were transduced with pCDH or pCDH-lncRNA- SNHG7 lentivirus followed by miR-34a-5p mimics or negative control transfection. Western blot analysis of LDHA and β-actin. (E, F) MCF-7 cells were transduced with sh-ctrl or sh-lncRNA- SNHG7 lentivirus followed by miR-34a-5p inhibitor or negative control transfection. ECAR was measured by Seahorse XF assays. Data were presented as mean ± standard deviation. lncRNA = long non-coding RNA; sh = short hairpin; ECAR = extracellular acidification rate; qRT-PCR = quantitative real-time polymerase chain reaction; ns = not significant; LDHA = lactate dehydrogenase A; 3′UTR = 3′ untranslated region. * p

    Article Snippet: Increased level of lncRNA-SNHG7 was detected in breast cancer tissue and cell lines To investigate the potential role of lncRNA-SNHG7 in breast cancer, we monitored the level of lncRNA-SNHG7 in breast cancer tissues.

    Techniques: Incubation, In Vitro, Synthesized, Labeling, Pull Down Assay, Quantitative RT-PCR, Transduction, Negative Control, Transfection, Western Blot, Standard Deviation, Real-time Polymerase Chain Reaction