qrt pcr analysis  (Roche)


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    Roche qrt pcr analysis
    Generating proliferative induced hepatocytes using defined transcription factors and oncogenic drivers ( A ) Schematic outline of the cell transformation assay for making lineage-specific cancer by lentiviral expression of three lineage-specific TFs to convert HFs to induced hepatocytes (iHep) and defined oncogenic drivers to transform iHeps to proliferating and tumorigenic cells. ( B ) Comparison of TF combinations ( Du et al , 2014 , Huang et al , 2014 , Morris et al , 2014 ) for converting human fibroblasts to iHeps by detecting transcript levels for liver marker genes ( ALBUMIN , TRANSFERRIN and SERPINA1/α-1-antitrypsin ) by <t>qRT-PCR</t> at different time points after iHep conversion, normalized to GAPDH levels (mean ± standard error).
    Qrt Pcr Analysis, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 296 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Cellular transformation by combined lineage conversion and oncogene expression"

    Article Title: Cellular transformation by combined lineage conversion and oncogene expression

    Journal: bioRxiv

    doi: 10.1101/525600

    Generating proliferative induced hepatocytes using defined transcription factors and oncogenic drivers ( A ) Schematic outline of the cell transformation assay for making lineage-specific cancer by lentiviral expression of three lineage-specific TFs to convert HFs to induced hepatocytes (iHep) and defined oncogenic drivers to transform iHeps to proliferating and tumorigenic cells. ( B ) Comparison of TF combinations ( Du et al , 2014 , Huang et al , 2014 , Morris et al , 2014 ) for converting human fibroblasts to iHeps by detecting transcript levels for liver marker genes ( ALBUMIN , TRANSFERRIN and SERPINA1/α-1-antitrypsin ) by qRT-PCR at different time points after iHep conversion, normalized to GAPDH levels (mean ± standard error).
    Figure Legend Snippet: Generating proliferative induced hepatocytes using defined transcription factors and oncogenic drivers ( A ) Schematic outline of the cell transformation assay for making lineage-specific cancer by lentiviral expression of three lineage-specific TFs to convert HFs to induced hepatocytes (iHep) and defined oncogenic drivers to transform iHeps to proliferating and tumorigenic cells. ( B ) Comparison of TF combinations ( Du et al , 2014 , Huang et al , 2014 , Morris et al , 2014 ) for converting human fibroblasts to iHeps by detecting transcript levels for liver marker genes ( ALBUMIN , TRANSFERRIN and SERPINA1/α-1-antitrypsin ) by qRT-PCR at different time points after iHep conversion, normalized to GAPDH levels (mean ± standard error).

    Techniques Used: Cell Transformation Assay, Expressing, Marker, Quantitative RT-PCR

    2) Product Images from "A Myosin-7B dependent endocytosis pathway mediates cellular entry of α-Synuclein fibrils and polycation-bearing cargos"

    Article Title: A Myosin-7B dependent endocytosis pathway mediates cellular entry of α-Synuclein fibrils and polycation-bearing cargos

    Journal: bioRxiv

    doi: 10.1101/2020.02.12.946137

    SLC35B2 is required for endocytosis of α-Syn PFF (A) The CRISPR screen strategy. (B-D) SLC35B2 -KO cells are defective in endocytosis of α-Syn PFF but not monomeric α-Syn. (B) Validation of SLC35B2 (35B2)-KO cells by qRT-PCR. mRNA levels were normalized to that in control cells. Error bar, SEM, n=3. (C) Control, SLC35B2 -KO, or SLC35B2 -KO cells stably expressing FLAG-SLC35B2 were incubated with pHrodo-labeled α-Syn PFF (200nM for 4 h) and analyzed by FACS. FL, fluorescence. (D) Control and SLC35B2 -KO cells were treated with labeled α-Syn monomer at 800nM overnight prior to FACS analysis. (E-G) Knockdown (KD) of SLC35B2 attenuates α-Syn PFF uptake in primary neurons. (E) Primary neurons were infected with lentivirus expressing the indicated shRNAs together with EGFP (driven by the Synapsin promoter) at days-in- vitro 3 (DIV3). mRNA was purified from cells at DIV8 for qRT-PCR analysis. Error bar, SEM, n=2 biological repeats, each with triplicated PCR analyses. (F) Control or SLC35B2 -KD neurons expressing EGFP (green) at DIV8 were incubated with α-Syn PFF Alexa 594 (200nM) (red) for 4 h, stained with DAPI (blue), and analyzed by confocal microscopy. Scale bar, 5µm (G) Quantification of α-Syn PFF level in individual cells (indicated by dots) from two independent experiments. Error bar, SEM, p value from two-tailed t-test. A.U. arbitrary unit. (H, I) SLC35B2 is required for α-Syn PFF binding to the plasma membrane. ( H ) Control, SLC35B2 -KO, or SLC35B2 -KO cells re-expressing WT SLC35B2 were incubated with α-Syn PFF Alexa 594 (200nM) (magenta) on ice for 30min, stained with DAPI (blue), and imaged by confocal microscopy. Scale bar, 5µm. The graph in (I) shows the quantification of α-Syn PFF surface level in individual cells from two independent experiments. A.U. arbitrary unit.
    Figure Legend Snippet: SLC35B2 is required for endocytosis of α-Syn PFF (A) The CRISPR screen strategy. (B-D) SLC35B2 -KO cells are defective in endocytosis of α-Syn PFF but not monomeric α-Syn. (B) Validation of SLC35B2 (35B2)-KO cells by qRT-PCR. mRNA levels were normalized to that in control cells. Error bar, SEM, n=3. (C) Control, SLC35B2 -KO, or SLC35B2 -KO cells stably expressing FLAG-SLC35B2 were incubated with pHrodo-labeled α-Syn PFF (200nM for 4 h) and analyzed by FACS. FL, fluorescence. (D) Control and SLC35B2 -KO cells were treated with labeled α-Syn monomer at 800nM overnight prior to FACS analysis. (E-G) Knockdown (KD) of SLC35B2 attenuates α-Syn PFF uptake in primary neurons. (E) Primary neurons were infected with lentivirus expressing the indicated shRNAs together with EGFP (driven by the Synapsin promoter) at days-in- vitro 3 (DIV3). mRNA was purified from cells at DIV8 for qRT-PCR analysis. Error bar, SEM, n=2 biological repeats, each with triplicated PCR analyses. (F) Control or SLC35B2 -KD neurons expressing EGFP (green) at DIV8 were incubated with α-Syn PFF Alexa 594 (200nM) (red) for 4 h, stained with DAPI (blue), and analyzed by confocal microscopy. Scale bar, 5µm (G) Quantification of α-Syn PFF level in individual cells (indicated by dots) from two independent experiments. Error bar, SEM, p value from two-tailed t-test. A.U. arbitrary unit. (H, I) SLC35B2 is required for α-Syn PFF binding to the plasma membrane. ( H ) Control, SLC35B2 -KO, or SLC35B2 -KO cells re-expressing WT SLC35B2 were incubated with α-Syn PFF Alexa 594 (200nM) (magenta) on ice for 30min, stained with DAPI (blue), and imaged by confocal microscopy. Scale bar, 5µm. The graph in (I) shows the quantification of α-Syn PFF surface level in individual cells from two independent experiments. A.U. arbitrary unit.

    Techniques Used: CRISPR, Quantitative RT-PCR, Stable Transfection, Expressing, Incubation, Labeling, FACS, Fluorescence, Infection, In Vitro, Purification, Polymerase Chain Reaction, Staining, Confocal Microscopy, Two Tailed Test, Binding Assay

    HSPG-mediated endocytosis requires MYO7B and actin filaments (A) Mapping the identified MYO7B sgRNA. (B) Validation of MYO7B -KO cells by qRT-PCR. mRNAs extracted from two clones of MYO7B KO cells and a control (Ctrl.) clone were analyzed by qRT-PCR. Error bars, SEM, n=3. (C) MYO7B KO cells are defective in α-Syn PFF uptake. Control (Ctrl.) and MYO7B -KO clones were incubated with pHrodo-labeled α-Syn PFF for 4 h before FACS analyses. (D) MYO7B is not required for endocytosis of α-Syn monomer. Control or MYO7B -KO cells were incubated with α-Syn monomer (800nM) overnight prior to FACS analysis. (E) MYO7B is not required for transferrin uptake. Control or MYO7B -KO cells were incubated with fluorescein-labeled transferrin (50µg/ml, 3h), washed, and analyzed with a fluorometer. Error bars, SEM, n=3. (F-H) KD of MYO7B in primary neurons reduces α-Syn PFF endocytosis without affecting its binding to the plasma membrane. (F) Primary neurons infected with the indicated shRNA-expressing lentivirus together with Synapsin_EGFP lentivirus at days-in- vitro 3 (DIV3) were incubated with α-Syn PFF Alexa 594 (200nM) for 4 h and imaged at DIV8. The insets show an enlarged view of the box. Arrows indicate peri-nuclear enrichment of α-Syn PFF positive vesicles in control cells. The graph in (G) shows the ratio of internalized α-Syn PFF relative to total α-Syn PFF in individual cells. (H) qRT-PCR evaluation of MYO7B mRNA levels using the same batch of cells. Error bars, SEM, n=two biological repeats with triplicated PCR analyses. (I-K) MYO7B dominant negative mutants inhibit α-Syn PFF uptake. (I) Representative confocal images of COS7 cells transfected with the indicated MYO7B plasmids and treated with α-Syn PFF Alexa 594 (400nM) for 3 h. Arrows indicate normal α-Syn PFF uptake in untransfected cells, which was used to normalize uptake. N, nuclei. (J) The graph shows the domain structure of the truncated MYO7B mutants. (K) Quantification of α-Syn PFF fluorescence (FL) in individual cells transfected (t) with the indicated MYO7B mutants normalized by the signal in untransfected cells (u).
    Figure Legend Snippet: HSPG-mediated endocytosis requires MYO7B and actin filaments (A) Mapping the identified MYO7B sgRNA. (B) Validation of MYO7B -KO cells by qRT-PCR. mRNAs extracted from two clones of MYO7B KO cells and a control (Ctrl.) clone were analyzed by qRT-PCR. Error bars, SEM, n=3. (C) MYO7B KO cells are defective in α-Syn PFF uptake. Control (Ctrl.) and MYO7B -KO clones were incubated with pHrodo-labeled α-Syn PFF for 4 h before FACS analyses. (D) MYO7B is not required for endocytosis of α-Syn monomer. Control or MYO7B -KO cells were incubated with α-Syn monomer (800nM) overnight prior to FACS analysis. (E) MYO7B is not required for transferrin uptake. Control or MYO7B -KO cells were incubated with fluorescein-labeled transferrin (50µg/ml, 3h), washed, and analyzed with a fluorometer. Error bars, SEM, n=3. (F-H) KD of MYO7B in primary neurons reduces α-Syn PFF endocytosis without affecting its binding to the plasma membrane. (F) Primary neurons infected with the indicated shRNA-expressing lentivirus together with Synapsin_EGFP lentivirus at days-in- vitro 3 (DIV3) were incubated with α-Syn PFF Alexa 594 (200nM) for 4 h and imaged at DIV8. The insets show an enlarged view of the box. Arrows indicate peri-nuclear enrichment of α-Syn PFF positive vesicles in control cells. The graph in (G) shows the ratio of internalized α-Syn PFF relative to total α-Syn PFF in individual cells. (H) qRT-PCR evaluation of MYO7B mRNA levels using the same batch of cells. Error bars, SEM, n=two biological repeats with triplicated PCR analyses. (I-K) MYO7B dominant negative mutants inhibit α-Syn PFF uptake. (I) Representative confocal images of COS7 cells transfected with the indicated MYO7B plasmids and treated with α-Syn PFF Alexa 594 (400nM) for 3 h. Arrows indicate normal α-Syn PFF uptake in untransfected cells, which was used to normalize uptake. N, nuclei. (J) The graph shows the domain structure of the truncated MYO7B mutants. (K) Quantification of α-Syn PFF fluorescence (FL) in individual cells transfected (t) with the indicated MYO7B mutants normalized by the signal in untransfected cells (u).

    Techniques Used: Quantitative RT-PCR, Clone Assay, Incubation, Labeling, FACS, Binding Assay, Infection, shRNA, Expressing, In Vitro, Polymerase Chain Reaction, Dominant Negative Mutation, Transfection, Fluorescence

    3) Product Images from "Antimicrobial peptides and nitric oxide production by neutrophils from periodontitis subjects"

    Article Title: Antimicrobial peptides and nitric oxide production by neutrophils from periodontitis subjects

    Journal: Brazilian Journal of Medical and Biological Research

    doi: 10.1590/S0100-879X2012007500123

    HNP 1-3 and LL-37 mRNA expression in peripheral neutrophils from periodontitis and healthy subjects. Neutrophils were isolated from peripheral blood and analyzed by flow cytometry, showing more than 94% purity according to side scatter (SSC) vs forward scatter (FSC) parameters ( A ). Neutrophils were incubated for 6 and 12 h with culture medium only or with medium containing Aa -LPS (100 ng/mL), Pg -LPS (100 ng/mL) or Ec -LPS (100 ng/mL). The relative mRNA expression of HNP 1-3 ( B ) and LL-37 ( C ) in neutrophils was assessed by qRT-PCR. Data are reported as means ± SD for N = 6 in each group. Aa -LPS = Aggregatibacter actinomycetemcomitans -lipopolysaccharide; Pg -LPS = Porphyromonas gingivalis -LPS; Ec -LPS = Escherichia coli -LPS. * P ≤ 0.05, periodontitis compared to healthy subjects (Student t -test).
    Figure Legend Snippet: HNP 1-3 and LL-37 mRNA expression in peripheral neutrophils from periodontitis and healthy subjects. Neutrophils were isolated from peripheral blood and analyzed by flow cytometry, showing more than 94% purity according to side scatter (SSC) vs forward scatter (FSC) parameters ( A ). Neutrophils were incubated for 6 and 12 h with culture medium only or with medium containing Aa -LPS (100 ng/mL), Pg -LPS (100 ng/mL) or Ec -LPS (100 ng/mL). The relative mRNA expression of HNP 1-3 ( B ) and LL-37 ( C ) in neutrophils was assessed by qRT-PCR. Data are reported as means ± SD for N = 6 in each group. Aa -LPS = Aggregatibacter actinomycetemcomitans -lipopolysaccharide; Pg -LPS = Porphyromonas gingivalis -LPS; Ec -LPS = Escherichia coli -LPS. * P ≤ 0.05, periodontitis compared to healthy subjects (Student t -test).

    Techniques Used: Expressing, Isolation, Flow Cytometry, Cytometry, Incubation, Quantitative RT-PCR

    4) Product Images from "JunB regulates homeostasis and suppressive functions of effector regulatory T cells"

    Article Title: JunB regulates homeostasis and suppressive functions of effector regulatory T cells

    Journal: Nature Communications

    doi: 10.1038/s41467-018-07735-4

    Expression of JunB is upregulated in eTreg cells. a – d Flow cytometry analysis of JunB in Foxp3 + (Treg) or Foxp3 − (Tconv) cells isolated from spleen a and lung b , Treg cells bearing CD62L hi CD44 lo phenotypes (cTreg) or CD62L lo phenotypes (eTreg) c , and ICOS + or ICOS − eTreg cells d isolated from spleen of wild-type C57BL/6 mice (7–10-week-old). Cd4 cre Junb fl/fl mice were used as negative controls. e CD62L hi CD44 lo cTreg and CD62L lo eTreg cells were sorted by FACS, and Junb mRNA expression was analyzed by qRT-PCR. a – e Error bars indicate s.d. ( n = 4). * P
    Figure Legend Snippet: Expression of JunB is upregulated in eTreg cells. a – d Flow cytometry analysis of JunB in Foxp3 + (Treg) or Foxp3 − (Tconv) cells isolated from spleen a and lung b , Treg cells bearing CD62L hi CD44 lo phenotypes (cTreg) or CD62L lo phenotypes (eTreg) c , and ICOS + or ICOS − eTreg cells d isolated from spleen of wild-type C57BL/6 mice (7–10-week-old). Cd4 cre Junb fl/fl mice were used as negative controls. e CD62L hi CD44 lo cTreg and CD62L lo eTreg cells were sorted by FACS, and Junb mRNA expression was analyzed by qRT-PCR. a – e Error bars indicate s.d. ( n = 4). * P

    Techniques Used: Expressing, Flow Cytometry, Cytometry, Isolation, Mouse Assay, FACS, Quantitative RT-PCR

    5) Product Images from "Pea p68, a DEAD-box helicase, enhances salt tolerance in marker-free transgenic plants of soybean [Glycine max (L.) Merrill]"

    Article Title: Pea p68, a DEAD-box helicase, enhances salt tolerance in marker-free transgenic plants of soybean [Glycine max (L.) Merrill]

    Journal: 3 Biotech

    doi: 10.1007/s13205-018-1553-z

    Expression analysis of transformed soybean plants. a RT-PCR expression analysis of p68 gene in transformed soybean plants. Lane 1, non-transformed (NT) soybean plant RNA as a negative control; lanes 2, 4 and 6, transformed (T 1 ) soybean (S1, S12 and S15) plant RNA samples; lanes 3 and 5, failed to amplify p68 gene from transformed (T 1 ) soybean (S2 and S13) plant RNA samples. b qRT-PCR analysis to analyze the expression level of p68 gene in transformed soybean plants. Relative expression of three transgenic lines (S1, S12 and S15) and non-transformed (NT) soybean plant; data were analyzed according to the 2 − ΔΔCt method. Mean of three individual experiments with standard errors. Different letters denote significantly different values according to Duncan’s multiple range test (DMRT) at a 5% level
    Figure Legend Snippet: Expression analysis of transformed soybean plants. a RT-PCR expression analysis of p68 gene in transformed soybean plants. Lane 1, non-transformed (NT) soybean plant RNA as a negative control; lanes 2, 4 and 6, transformed (T 1 ) soybean (S1, S12 and S15) plant RNA samples; lanes 3 and 5, failed to amplify p68 gene from transformed (T 1 ) soybean (S2 and S13) plant RNA samples. b qRT-PCR analysis to analyze the expression level of p68 gene in transformed soybean plants. Relative expression of three transgenic lines (S1, S12 and S15) and non-transformed (NT) soybean plant; data were analyzed according to the 2 − ΔΔCt method. Mean of three individual experiments with standard errors. Different letters denote significantly different values according to Duncan’s multiple range test (DMRT) at a 5% level

    Techniques Used: Expressing, Transformation Assay, Reverse Transcription Polymerase Chain Reaction, Negative Control, Quantitative RT-PCR, Transgenic Assay

    6) Product Images from "Danger-Associated Peptide Regulates Root Immune Responses and Root Growth by Affecting ROS Formation in Arabidopsis"

    Article Title: Danger-Associated Peptide Regulates Root Immune Responses and Root Growth by Affecting ROS Formation in Arabidopsis

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21134590

    BIK1 is required to regulate Pep1-induced ROS production and root growth inhibition. ( A ) Histochemical staining of GUS activity in the roots of transgenic plants harboring proBIK1:GUS upon exposure to Pep1. The six-day-old plants were transferred on half-strength MS agar medium, supplemented with 1 μM Pep1 for 1, 2, 4, 6, 8, 10, and 12 h, respectively. The experiments were repeated three times with similar observations. Bars = 200 μm. ( B ) qRT-PCR analysis of mRNA levels of BIK1 in the wild type (WT) roots. The two-week-old plants were incubated in half-strength MS liquid medium, supplemented with 1 μM Pep1 for 0.5, 1, 2, 4, 8, 12, and 24 h, respectively. The expression level of control (0 h) was set as 1.0 and the Pep1 treatment levels were normalized to the control level. Data are means ± SD ( n = 3). ( C ) H 2 DCF-DA staining of H 2 O 2 in WT, bik1 , 35S:BIK1 / bik1, pepr1 bik1 , pepr2 bik1, and pepr1 pepr2 bik1 roots. Six-day-old seedlings were incubated in half-strength MS liquid medium, supplemented with 1 μM Pep1 for 18 h. The roots were stained with 30 μM H 2 DCF-DA solution for 10 min and photographed under fluorescence microscopy (Olympus, BX53). Bars = 100 μm. ( D ) The statistical analysis of ROS production, as indicated in ( C ). Data are means ± SD from three independent experiments. ( n = 30). The relative ROS production of each treatment was normalized to control of wild type roots (100%). ( E ) The growth phenotype of WT, bik1 , 35S:BIK1 / bik1, pepr1 bik1 , pepr2 bik1, and pepr1 pepr2 bik1 roots. The three-day-old seedlings were transplanted on half-strength MS agar medium supplemented with or without (control) 100 nM Pep1 for six days. ( F ) The statistical analysis of the primary root length, as indicated in ( E ). Data are means ± SD from three independent experiments ( n = 15). Asterisks in ( B , D , F ) indicate statistically significant differences compared with the untreated controls in each genotype (Tukey’s test, * p
    Figure Legend Snippet: BIK1 is required to regulate Pep1-induced ROS production and root growth inhibition. ( A ) Histochemical staining of GUS activity in the roots of transgenic plants harboring proBIK1:GUS upon exposure to Pep1. The six-day-old plants were transferred on half-strength MS agar medium, supplemented with 1 μM Pep1 for 1, 2, 4, 6, 8, 10, and 12 h, respectively. The experiments were repeated three times with similar observations. Bars = 200 μm. ( B ) qRT-PCR analysis of mRNA levels of BIK1 in the wild type (WT) roots. The two-week-old plants were incubated in half-strength MS liquid medium, supplemented with 1 μM Pep1 for 0.5, 1, 2, 4, 8, 12, and 24 h, respectively. The expression level of control (0 h) was set as 1.0 and the Pep1 treatment levels were normalized to the control level. Data are means ± SD ( n = 3). ( C ) H 2 DCF-DA staining of H 2 O 2 in WT, bik1 , 35S:BIK1 / bik1, pepr1 bik1 , pepr2 bik1, and pepr1 pepr2 bik1 roots. Six-day-old seedlings were incubated in half-strength MS liquid medium, supplemented with 1 μM Pep1 for 18 h. The roots were stained with 30 μM H 2 DCF-DA solution for 10 min and photographed under fluorescence microscopy (Olympus, BX53). Bars = 100 μm. ( D ) The statistical analysis of ROS production, as indicated in ( C ). Data are means ± SD from three independent experiments. ( n = 30). The relative ROS production of each treatment was normalized to control of wild type roots (100%). ( E ) The growth phenotype of WT, bik1 , 35S:BIK1 / bik1, pepr1 bik1 , pepr2 bik1, and pepr1 pepr2 bik1 roots. The three-day-old seedlings were transplanted on half-strength MS agar medium supplemented with or without (control) 100 nM Pep1 for six days. ( F ) The statistical analysis of the primary root length, as indicated in ( E ). Data are means ± SD from three independent experiments ( n = 15). Asterisks in ( B , D , F ) indicate statistically significant differences compared with the untreated controls in each genotype (Tukey’s test, * p

    Techniques Used: Inhibition, Staining, Activity Assay, Transgenic Assay, Quantitative RT-PCR, Incubation, Expressing, Fluorescence, Microscopy

    7) Product Images from "Upregulation of erythropoietin and erythropoietin receptor in castration-resistant progression of prostate cancer"

    Article Title: Upregulation of erythropoietin and erythropoietin receptor in castration-resistant progression of prostate cancer

    Journal: Asian Journal of Andrology

    doi: 10.4103/aja.aja_80_19

    ( a ) Expression levels of EPO and EPOR in an LNCaP cell culture under hypoxic conditions and normoxic conditions with CS-FBS, as determined by qRT-PCR. ( b ) Expression levels of EPO and EPOR in hypoxia-induced LNCaP cells and LNCaP cells under normoxic condition or CS-FBS, as determined by western blot. The quantification analysis is presented in the right panel. ( c ) EPOR knockdown using siRNA, as determined by qRT-PCR. ( d ) The expression of NSE in LNCaP cells cultured under hypoxic conditions after EPOR knockdown, determined by western blot. The quantification analysis is presented in the right panel. ( e ) Cell proliferation results of LNCap cells cultured under hypoxic conditions in a CS-FBS culture and normoxic conditions with a complete medium after EPOR knockdown, as determined by a CCK8 assay. The experiments were performed in triplicate * P > 0.05, ** P
    Figure Legend Snippet: ( a ) Expression levels of EPO and EPOR in an LNCaP cell culture under hypoxic conditions and normoxic conditions with CS-FBS, as determined by qRT-PCR. ( b ) Expression levels of EPO and EPOR in hypoxia-induced LNCaP cells and LNCaP cells under normoxic condition or CS-FBS, as determined by western blot. The quantification analysis is presented in the right panel. ( c ) EPOR knockdown using siRNA, as determined by qRT-PCR. ( d ) The expression of NSE in LNCaP cells cultured under hypoxic conditions after EPOR knockdown, determined by western blot. The quantification analysis is presented in the right panel. ( e ) Cell proliferation results of LNCap cells cultured under hypoxic conditions in a CS-FBS culture and normoxic conditions with a complete medium after EPOR knockdown, as determined by a CCK8 assay. The experiments were performed in triplicate * P > 0.05, ** P

    Techniques Used: Expressing, Cell Culture, Quantitative RT-PCR, Western Blot, CCK-8 Assay

    8) Product Images from "A member of the ETS family, EHF, and the ATPase RUVBL1 inhibit p53-mediated apoptosis"

    Article Title: A member of the ETS family, EHF, and the ATPase RUVBL1 inhibit p53-mediated apoptosis

    Journal: EMBO Reports

    doi: 10.1038/embor.2011.81

    EHF directly regulates the expression of RUVBL1. ( A ) qRT–PCR analysis of RUVBL1 expression in HCT116 cells transfected with siRNA targeting a scrambled sequence (siCont) or EHF (siEHF1, siEHF2). Before fold-change calculation, values were normalized
    Figure Legend Snippet: EHF directly regulates the expression of RUVBL1. ( A ) qRT–PCR analysis of RUVBL1 expression in HCT116 cells transfected with siRNA targeting a scrambled sequence (siCont) or EHF (siEHF1, siEHF2). Before fold-change calculation, values were normalized

    Techniques Used: Expressing, Quantitative RT-PCR, Transfection, Sequencing

    9) Product Images from "The novel lncRNA lnc-NR2F1 is pro-neurogenic and mutated in human neurodevelopmental disorders"

    Article Title: The novel lncRNA lnc-NR2F1 is pro-neurogenic and mutated in human neurodevelopmental disorders

    Journal: eLife

    doi: 10.7554/eLife.41770

    Characterization of the epistasis relationship between mouse Nr2f1 and lnc-Nr2f1 . ( A ) CRISPR knock out strategy to generate Nr2f1 knockout (Homo) and heterozygous null lines (Het) from the control mES cells (Ctrl). ( B ) qRT-PCR results for lnc-Nr2f1 and Nr2f1 in the Ctrl, Nr2f1 heterozygous null and Nr2f1 knock out day 4 Ngn2 mES-iN. (n = 4 for Ctrl and Homo, n = 6 for Het; n.s. denotes not significant by two tailed t test) ( C ) Western blot showing the level of NR2F1 for individual clones of Ctrl, Het and Homo for day 4 Ngn2 mES-iN. ( D ) Neurite length measurement of the Ngn2 day 3 mES iN cells generated from the Nr2f1 Ctrl, Het or Homo lines. (n = 4 for Ctrl and Homo, n = 6 for Het) (n.s. indicates p
    Figure Legend Snippet: Characterization of the epistasis relationship between mouse Nr2f1 and lnc-Nr2f1 . ( A ) CRISPR knock out strategy to generate Nr2f1 knockout (Homo) and heterozygous null lines (Het) from the control mES cells (Ctrl). ( B ) qRT-PCR results for lnc-Nr2f1 and Nr2f1 in the Ctrl, Nr2f1 heterozygous null and Nr2f1 knock out day 4 Ngn2 mES-iN. (n = 4 for Ctrl and Homo, n = 6 for Het; n.s. denotes not significant by two tailed t test) ( C ) Western blot showing the level of NR2F1 for individual clones of Ctrl, Het and Homo for day 4 Ngn2 mES-iN. ( D ) Neurite length measurement of the Ngn2 day 3 mES iN cells generated from the Nr2f1 Ctrl, Het or Homo lines. (n = 4 for Ctrl and Homo, n = 6 for Het) (n.s. indicates p

    Techniques Used: CRISPR, Knock-Out, Quantitative RT-PCR, Two Tailed Test, Western Blot, Clone Assay, Generated

    QRT-PCR validation of candidate lncRNAs expression. ( A ) Expression detection of candidate lncRNAs by qRT-PCR across early stages of iN cell reprogramming and mouse brain development.
    Figure Legend Snippet: QRT-PCR validation of candidate lncRNAs expression. ( A ) Expression detection of candidate lncRNAs by qRT-PCR across early stages of iN cell reprogramming and mouse brain development.

    Techniques Used: Quantitative RT-PCR, Expressing

    10) Product Images from "Identification of novel MYB transcription factors involved in the isoflavone biosynthetic pathway by using the combination screening system with agroinfiltration and hairy root transformation"

    Article Title: Identification of novel MYB transcription factors involved in the isoflavone biosynthetic pathway by using the combination screening system with agroinfiltration and hairy root transformation

    Journal: Plant Biotechnology

    doi: 10.5511/plantbiotechnology.19.1025a

    Figure 4. Hierarchical clustering and heat map analysis of gene expression profiles and the accumulation of isoflavones in developing soybean seeds. The columns represent the expression profiles of individual MYB TFs, isoflavone biosynthetic genes, and isoflavone content in developing seeds. The rows represent seed developmental stages. The white-color box in the heat map indicated that out of three replicates the expression of gene could not be detected in all the replication in qRT-PCR. The highly correlated three MYB TFs, GmMYB102, GmMYB280, GmMYB502 with isoflavones content in developing seeds are indicated by red characters. The analysis was performed in R software.
    Figure Legend Snippet: Figure 4. Hierarchical clustering and heat map analysis of gene expression profiles and the accumulation of isoflavones in developing soybean seeds. The columns represent the expression profiles of individual MYB TFs, isoflavone biosynthetic genes, and isoflavone content in developing seeds. The rows represent seed developmental stages. The white-color box in the heat map indicated that out of three replicates the expression of gene could not be detected in all the replication in qRT-PCR. The highly correlated three MYB TFs, GmMYB102, GmMYB280, GmMYB502 with isoflavones content in developing seeds are indicated by red characters. The analysis was performed in R software.

    Techniques Used: Expressing, Quantitative RT-PCR, Software

    11) Product Images from "Knockdown of miR-572 suppresses cell proliferation and promotes apoptosis in renal cell carcinoma cells by targeting the NF2/Hippo signaling pathway"

    Article Title: Knockdown of miR-572 suppresses cell proliferation and promotes apoptosis in renal cell carcinoma cells by targeting the NF2/Hippo signaling pathway

    Journal: International Journal of Clinical and Experimental Pathology

    doi:

    miR-572 expression was upregulated in RCC specimens and cell lines.Relative miR-572 expression was assessed by qRT-PCR assay in RCC clinical specimens and adjacent non-cancerous specimens (A), and RCC cell lines (A498, 786-O, ACHN) and human renal tubule epithelial cell line HK-2 (B). * P
    Figure Legend Snippet: miR-572 expression was upregulated in RCC specimens and cell lines.Relative miR-572 expression was assessed by qRT-PCR assay in RCC clinical specimens and adjacent non-cancerous specimens (A), and RCC cell lines (A498, 786-O, ACHN) and human renal tubule epithelial cell line HK-2 (B). * P

    Techniques Used: Expressing, Quantitative RT-PCR

    Knockdown of miR-572 suppressed proliferation and enhanced apoptosis in RCC cells. 786-O cells were transfected with anti-miR-572 or anti-miR-NC, following the detection of miR-572 expression by qRT-PCR analysis (A), cell proliferation capability by CCK-8 assay (B), and cell apoptosis ability by flow cytometry (C). * P
    Figure Legend Snippet: Knockdown of miR-572 suppressed proliferation and enhanced apoptosis in RCC cells. 786-O cells were transfected with anti-miR-572 or anti-miR-NC, following the detection of miR-572 expression by qRT-PCR analysis (A), cell proliferation capability by CCK-8 assay (B), and cell apoptosis ability by flow cytometry (C). * P

    Techniques Used: Transfection, Expressing, Quantitative RT-PCR, CCK-8 Assay, Flow Cytometry, Cytometry

    Anti-miR-572-mediates the regulatory effect on proliferation and apoptosis of RCC cells. This was abated by the restoration of NF2 expression. 786-O cells were transfected with anti-miR-572 alone or together with si-NF2, following the measurement of relative NF2 expression by qRT-PCR (A), cell proliferation capacity by CCK-8 assay (B), and cell apoptosis ability by flow cytometry (C). * P
    Figure Legend Snippet: Anti-miR-572-mediates the regulatory effect on proliferation and apoptosis of RCC cells. This was abated by the restoration of NF2 expression. 786-O cells were transfected with anti-miR-572 alone or together with si-NF2, following the measurement of relative NF2 expression by qRT-PCR (A), cell proliferation capacity by CCK-8 assay (B), and cell apoptosis ability by flow cytometry (C). * P

    Techniques Used: Expressing, Transfection, Quantitative RT-PCR, CCK-8 Assay, Flow Cytometry, Cytometry

    12) Product Images from "Pea p68, a DEAD-box helicase, enhances salt tolerance in marker-free transgenic plants of soybean [Glycine max (L.) Merrill]"

    Article Title: Pea p68, a DEAD-box helicase, enhances salt tolerance in marker-free transgenic plants of soybean [Glycine max (L.) Merrill]

    Journal: 3 Biotech

    doi: 10.1007/s13205-018-1553-z

    Expression analysis of transformed soybean plants. a RT-PCR expression analysis of p68 gene in transformed soybean plants. Lane 1, non-transformed (NT) soybean plant RNA as a negative control; lanes 2, 4 and 6, transformed (T 1 ) soybean (S1, S12 and S15) plant RNA samples; lanes 3 and 5, failed to amplify p68 gene from transformed (T 1 ) soybean (S2 and S13) plant RNA samples. b qRT-PCR analysis to analyze the expression level of p68 gene in transformed soybean plants. Relative expression of three transgenic lines (S1, S12 and S15) and non-transformed (NT) soybean plant; data were analyzed according to the 2 − ΔΔCt method. Mean of three individual experiments with standard errors. Different letters denote significantly different values according to Duncan’s multiple range test (DMRT) at a 5% level
    Figure Legend Snippet: Expression analysis of transformed soybean plants. a RT-PCR expression analysis of p68 gene in transformed soybean plants. Lane 1, non-transformed (NT) soybean plant RNA as a negative control; lanes 2, 4 and 6, transformed (T 1 ) soybean (S1, S12 and S15) plant RNA samples; lanes 3 and 5, failed to amplify p68 gene from transformed (T 1 ) soybean (S2 and S13) plant RNA samples. b qRT-PCR analysis to analyze the expression level of p68 gene in transformed soybean plants. Relative expression of three transgenic lines (S1, S12 and S15) and non-transformed (NT) soybean plant; data were analyzed according to the 2 − ΔΔCt method. Mean of three individual experiments with standard errors. Different letters denote significantly different values according to Duncan’s multiple range test (DMRT) at a 5% level

    Techniques Used: Expressing, Transformation Assay, Reverse Transcription Polymerase Chain Reaction, Negative Control, Quantitative RT-PCR, Transgenic Assay

    13) Product Images from "Transcriptome Analysis Identifies Candidate Target Genes Involved in Glyphosate-Resistance Mechanism in Lolium multiflorum"

    Article Title: Transcriptome Analysis Identifies Candidate Target Genes Involved in Glyphosate-Resistance Mechanism in Lolium multiflorum

    Journal: Plants

    doi: 10.3390/plants9060685

    The correlation between transcriptomic (RNA-Seq; X-axis) and qRT-PCR (Y-axis) fold-change (GR/GS) expression levels of 19 genes of glyphosate-resistant (GR) and glyphosate-sensitive (GS) biotypes of Lolium multiflorum .
    Figure Legend Snippet: The correlation between transcriptomic (RNA-Seq; X-axis) and qRT-PCR (Y-axis) fold-change (GR/GS) expression levels of 19 genes of glyphosate-resistant (GR) and glyphosate-sensitive (GS) biotypes of Lolium multiflorum .

    Techniques Used: RNA Sequencing Assay, Quantitative RT-PCR, Expressing

    14) Product Images from "Targeting heparan sulfate proteoglycan-assisted endocytosis as a COVID-19 therapeutic option"

    Article Title: Targeting heparan sulfate proteoglycan-assisted endocytosis as a COVID-19 therapeutic option

    Journal: bioRxiv

    doi: 10.1101/2020.07.14.202549

    Establishing tools to study ACE2-mediated coronaviral entry. ( A, B ) The entry of SARS-CoV or SARS-CoV-2 PPs into Calu-3 cells. Uninfected Calu-3 cells or cells infected with the indicated PPs for 48 h were analyzed for luciferase expression. Uninfected cells were normalized to 1. Error bars indicate SEM. N=4. ( C, D ) The entry of SARS-CoV or SARS-CoV-2 PPs is dependent on ACE2. HEK293T cells or HEK293T-ACE2-GFP cells were either uninfected or infected with the indicated PPs for 24h. The luciferase/GFP ratio was determined to indicate viral entry. Error bars indicate SEM. N=4. ( E ) Verification of XYLT2 knockdown by qRT-PCR. A fraction of ACE2-GFP cells transfected with SMARTpooled XYLT2 siRNAs or a control siRNA for 72 h were analyzed for XYLT2 expression by qRT-PCR. The remaining cells were used in Figure 1F for viral entry and cell viability assay. Error bars indicate SEM, N=3. (F) Generating ACE2-GFP cells deficient for SLC35B2 ( 35B2 ). ACE2-GFP cells treated with SLC35B2 sgRNA-expressing lentiviruses were incubated with Alex 594 -labeled (400 nM) α-Syn fibrils for 4h. α-Syn negative SLC35B2 knockout (KO) cells (dashed box) were identified and collected by FACS. ( F ) Verification of SLC35B2 knockdown by qRT-PCR. A fraction of ACE2-GFP cells transfected with SMARTpooled SLC35B2 siRNAs or a control siRNA for 72 h were analyzed for SLC35B2 expression by qRT-PCR. The remaining cells were used for viral entry and cell viability assay in Figure 1G . Error bars indicate SEM, N=3. ( H ) Knockout of SCL35B2 does not affect ACE2-GFP expression. The ACE2-GFP level in cell extracts used in Figure 1G was determined by a fluorometer.
    Figure Legend Snippet: Establishing tools to study ACE2-mediated coronaviral entry. ( A, B ) The entry of SARS-CoV or SARS-CoV-2 PPs into Calu-3 cells. Uninfected Calu-3 cells or cells infected with the indicated PPs for 48 h were analyzed for luciferase expression. Uninfected cells were normalized to 1. Error bars indicate SEM. N=4. ( C, D ) The entry of SARS-CoV or SARS-CoV-2 PPs is dependent on ACE2. HEK293T cells or HEK293T-ACE2-GFP cells were either uninfected or infected with the indicated PPs for 24h. The luciferase/GFP ratio was determined to indicate viral entry. Error bars indicate SEM. N=4. ( E ) Verification of XYLT2 knockdown by qRT-PCR. A fraction of ACE2-GFP cells transfected with SMARTpooled XYLT2 siRNAs or a control siRNA for 72 h were analyzed for XYLT2 expression by qRT-PCR. The remaining cells were used in Figure 1F for viral entry and cell viability assay. Error bars indicate SEM, N=3. (F) Generating ACE2-GFP cells deficient for SLC35B2 ( 35B2 ). ACE2-GFP cells treated with SLC35B2 sgRNA-expressing lentiviruses were incubated with Alex 594 -labeled (400 nM) α-Syn fibrils for 4h. α-Syn negative SLC35B2 knockout (KO) cells (dashed box) were identified and collected by FACS. ( F ) Verification of SLC35B2 knockdown by qRT-PCR. A fraction of ACE2-GFP cells transfected with SMARTpooled SLC35B2 siRNAs or a control siRNA for 72 h were analyzed for SLC35B2 expression by qRT-PCR. The remaining cells were used for viral entry and cell viability assay in Figure 1G . Error bars indicate SEM, N=3. ( H ) Knockout of SCL35B2 does not affect ACE2-GFP expression. The ACE2-GFP level in cell extracts used in Figure 1G was determined by a fluorometer.

    Techniques Used: Infection, Luciferase, Expressing, Quantitative RT-PCR, Transfection, Viability Assay, Incubation, Labeling, Knock-Out, FACS

    15) Product Images from "Crx broadly modulates the pineal transcriptome"

    Article Title: Crx broadly modulates the pineal transcriptome

    Journal: Journal of neurochemistry

    doi: 10.1111/j.1471-4159.2011.07405.x

    qRT-PCR analysis of transcripts detected as either day/night expressed or differentially expressed in Crx −/− mice as compared with wild-types
    Figure Legend Snippet: qRT-PCR analysis of transcripts detected as either day/night expressed or differentially expressed in Crx −/− mice as compared with wild-types

    Techniques Used: Quantitative RT-PCR, Mouse Assay

    16) Product Images from "CRISPR/Cas9-mediated targeted mutagenesis of GmLHY genes alters plant height and internode length in soybean"

    Article Title: CRISPR/Cas9-mediated targeted mutagenesis of GmLHY genes alters plant height and internode length in soybean

    Journal: BMC Plant Biology

    doi: 10.1186/s12870-019-2145-8

    Diurnal expression patterns of GmLHY1a/1b/2a/2b in WT plants and T2 homozygous quadruple mutants of GmLHY . a – d qRT-PCR analysis of GmLHY2b , GmLHY2a, GmLHY1a, and GmLHY1b expression levels in the leaves at 20 DAE under 16 h light/8 h dark (long day; LD) conditions, respectively. Data shown are relative to the control gene GmTUB and represent means ± standard error of the mean (s.e.m.) for three biological replicates. Bars indicate the s.e.m. Black and white bars represent dark and light periods, respectively
    Figure Legend Snippet: Diurnal expression patterns of GmLHY1a/1b/2a/2b in WT plants and T2 homozygous quadruple mutants of GmLHY . a – d qRT-PCR analysis of GmLHY2b , GmLHY2a, GmLHY1a, and GmLHY1b expression levels in the leaves at 20 DAE under 16 h light/8 h dark (long day; LD) conditions, respectively. Data shown are relative to the control gene GmTUB and represent means ± standard error of the mean (s.e.m.) for three biological replicates. Bars indicate the s.e.m. Black and white bars represent dark and light periods, respectively

    Techniques Used: Expressing, Quantitative RT-PCR

    17) Product Images from "High-density array analysis of DNA methylation in Tamoxifen-resistant breast cancer cell lines"

    Article Title: High-density array analysis of DNA methylation in Tamoxifen-resistant breast cancer cell lines

    Journal: Epigenetics

    doi: 10.4161/epi.27111

    Figure 6. Comparison of gene expression and promoter methylation in ZNF350 and MAGED1. Relative mRNA levels measured by qRT-PCR and average percent methylation of the TSS200 regions measured by pyrosequencing of ( A ) ZNF350 and ( B ) MAGED1 in control
    Figure Legend Snippet: Figure 6. Comparison of gene expression and promoter methylation in ZNF350 and MAGED1. Relative mRNA levels measured by qRT-PCR and average percent methylation of the TSS200 regions measured by pyrosequencing of ( A ) ZNF350 and ( B ) MAGED1 in control

    Techniques Used: Expressing, Methylation, Quantitative RT-PCR

    18) Product Images from "MicroRNA-21 promotes cell proliferation by targeting tumor suppressor TET1 in colorectal cancer"

    Article Title: MicroRNA-21 promotes cell proliferation by targeting tumor suppressor TET1 in colorectal cancer

    Journal: International Journal of Clinical and Experimental Pathology

    doi:

    MiR-21 was able to down-regulate TET1 in CRC cells. A, B. The mRNA and protein levels of TET1 were examined in HCT15 and HT29 cells transfected with anti-miR-21 by qRT-PCR and Western blotting, respectively. Statistically significant differences are indicated: * P
    Figure Legend Snippet: MiR-21 was able to down-regulate TET1 in CRC cells. A, B. The mRNA and protein levels of TET1 were examined in HCT15 and HT29 cells transfected with anti-miR-21 by qRT-PCR and Western blotting, respectively. Statistically significant differences are indicated: * P

    Techniques Used: Transfection, Quantitative RT-PCR, Western Blot

    MiR-21 was negatively associated with TET1 in clinical CRC tissues. A. TET1 mRNA levels was examined by qRT-PCR in 50 cases of clinical CRC tissues and paired adjacent tissues. B. Correlation of MiR-21 levels with TET1 mRNA levels was examined by qRT-PCR in 50 cases of clinical CRC tissues (Pearson’s correlation coefficient, r = -0.409). Statistically significant differences are indicated: * P
    Figure Legend Snippet: MiR-21 was negatively associated with TET1 in clinical CRC tissues. A. TET1 mRNA levels was examined by qRT-PCR in 50 cases of clinical CRC tissues and paired adjacent tissues. B. Correlation of MiR-21 levels with TET1 mRNA levels was examined by qRT-PCR in 50 cases of clinical CRC tissues (Pearson’s correlation coefficient, r = -0.409). Statistically significant differences are indicated: * P

    Techniques Used: Quantitative RT-PCR

    19) Product Images from "Effect of Sodium Butyrate on LHX1 mRNA Expression as a Transcription Factor of HDAC8 in Human Colorectal Cancer Cell Lines"

    Article Title: Effect of Sodium Butyrate on LHX1 mRNA Expression as a Transcription Factor of HDAC8 in Human Colorectal Cancer Cell Lines

    Journal: Avicenna Journal of Medical Biotechnology

    doi:

    The Effect of sodium butyrate (SB) on LHX1 mRNA expression in HCT-116 cell line. A) Cells were cultured for 24 hr with 6.25 mM to 100 mM of SB at 37 °C . B) Cells were cultured for 48 hr with 6.25 mM 100 mM of SB at 37 °C . LHX1 mRNA expression investigated using qRT-PCR. GAPDH was used as an internal control. LHX1 mRNA expression decreased in treated cells compared to control (0 mM ). * Indicates a significant reduction (p
    Figure Legend Snippet: The Effect of sodium butyrate (SB) on LHX1 mRNA expression in HCT-116 cell line. A) Cells were cultured for 24 hr with 6.25 mM to 100 mM of SB at 37 °C . B) Cells were cultured for 48 hr with 6.25 mM 100 mM of SB at 37 °C . LHX1 mRNA expression investigated using qRT-PCR. GAPDH was used as an internal control. LHX1 mRNA expression decreased in treated cells compared to control (0 mM ). * Indicates a significant reduction (p

    Techniques Used: Expressing, Cell Culture, Quantitative RT-PCR

    The effect of sodium butyrate (SB) on the LHX1 mRNA expression in HT-29 cell line. A) Cells were cultured for 24 hr with 6.25 to 100 mM concentrations of SB at 37 °C . B) Cells were cultured for 48 hr with 6.25 to 100 mM concentrations of SB at 37 °C . LHX1 mRNA expression was investigated using qRT-PCR. GAPDH was used as the internal control. LHX1 mRNA expression increased in treated cells compared to control (0 mM ). * Indicates a significant increase (p
    Figure Legend Snippet: The effect of sodium butyrate (SB) on the LHX1 mRNA expression in HT-29 cell line. A) Cells were cultured for 24 hr with 6.25 to 100 mM concentrations of SB at 37 °C . B) Cells were cultured for 48 hr with 6.25 to 100 mM concentrations of SB at 37 °C . LHX1 mRNA expression was investigated using qRT-PCR. GAPDH was used as the internal control. LHX1 mRNA expression increased in treated cells compared to control (0 mM ). * Indicates a significant increase (p

    Techniques Used: Expressing, Cell Culture, Quantitative RT-PCR

    20) Product Images from "Helicobacter pylori Vacuolating Cytotoxin A Causes Anorexia and Anxiety via Hypothalamic Urocortin 1 in Mice"

    Article Title: Helicobacter pylori Vacuolating Cytotoxin A Causes Anorexia and Anxiety via Hypothalamic Urocortin 1 in Mice

    Journal: Scientific Reports

    doi: 10.1038/s41598-019-42163-4

    Peripheral VacA administration induces anorexia, anxiety, and the mRNA expression of Ucn1 in mice. ( a) Cumulative food intake was measured for 24 h in mice receiving IP administration ( n = 8–10). ( b , c ) The percentage of time spent in the open arms ( b ) and total distance ( c ) were measured 4 h after the IP administration of 30 nmol/kg bw of VacA in mice ( n = 5–8). TBS (vehicle) was administered as the control. ( d ) Four hours after IP administration of 30 nmol/kg bw of VacA to mice, the orexigenic and anorexigenic peptide mRNA levels in the hypothalamus were measured by qRT-PCR ( n = 5–8). The values are presented as the means ± SEM. Differences were considered significant at * p
    Figure Legend Snippet: Peripheral VacA administration induces anorexia, anxiety, and the mRNA expression of Ucn1 in mice. ( a) Cumulative food intake was measured for 24 h in mice receiving IP administration ( n = 8–10). ( b , c ) The percentage of time spent in the open arms ( b ) and total distance ( c ) were measured 4 h after the IP administration of 30 nmol/kg bw of VacA in mice ( n = 5–8). TBS (vehicle) was administered as the control. ( d ) Four hours after IP administration of 30 nmol/kg bw of VacA to mice, the orexigenic and anorexigenic peptide mRNA levels in the hypothalamus were measured by qRT-PCR ( n = 5–8). The values are presented as the means ± SEM. Differences were considered significant at * p

    Techniques Used: Expressing, Mouse Assay, Quantitative RT-PCR

    21) Product Images from "Endogenous DUX4 expression in FSHD myotubes is sufficient to cause cell death and disrupts RNA splicing and cell migration pathways"

    Article Title: Endogenous DUX4 expression in FSHD myotubes is sufficient to cause cell death and disrupts RNA splicing and cell migration pathways

    Journal: Human Molecular Genetics

    doi: 10.1093/hmg/ddv315

    Quantification of DUX4 activation events and DUX4 and DUX4 target mRNA expression analysis in sorted myoblasts. ( A ) Representative flow cytometry plots of BFP fluorescence intensity (X-axis) versus autofluorescence (Y-axis) in 72 h differentiated control and FSHD myoblasts harboring the DUX4-activated BFP reporter with % reporter + cells displayed for each line. ( B ) Populations of BFP+ and BFP– myoblasts were collected by flow sorting and mRNA levels of DUX4, and DUX4 targets BFP, CCNA1 and MBD3L2 were measured by qRT-PCR. (N.D., not detected).
    Figure Legend Snippet: Quantification of DUX4 activation events and DUX4 and DUX4 target mRNA expression analysis in sorted myoblasts. ( A ) Representative flow cytometry plots of BFP fluorescence intensity (X-axis) versus autofluorescence (Y-axis) in 72 h differentiated control and FSHD myoblasts harboring the DUX4-activated BFP reporter with % reporter + cells displayed for each line. ( B ) Populations of BFP+ and BFP– myoblasts were collected by flow sorting and mRNA levels of DUX4, and DUX4 targets BFP, CCNA1 and MBD3L2 were measured by qRT-PCR. (N.D., not detected).

    Techniques Used: Activation Assay, Expressing, Flow Cytometry, Cytometry, Fluorescence, Quantitative RT-PCR

    Characterization of a DUX4-activated fluorescent reporter. ( A ) A schematic diagram of the lentiviral vector encoding a DUX4 reporter with five unique DUX4 binding site sequences, identified from individual DUX4 genomic targets and a TATA box located upstream of the sequence for nuclear-localized Blue Fluorescent Protein (nuBFP). A pTK and neomycin phosphotransferase gene (NEO) were included to allow for selection of transduced cells. ( B ) BFP is specifically present in MHC-positive myotubes (green) derived from individuals with FSHD and co-localizes with DUX4 protein (red) by immunofluorescence. ( C ) Delivery of an siRNA targeting the DUX4 transcript (siDUX4) during FSHD myoblast differentiation prohibits BFP reporter activation by eliminating DUX4 protein expression. A universal non-targeting control siRNA (siCTRL) transfected in parallel results in BFP and DUX4 expression typical of FSHD myotubes. ( D ) qRT-PCR of DUX4 and DUX4 targets BFP, CCNA1 and MBD3L2 shows reduced mRNA levels after siDUX4 delivery during FSHD myoblast differentiation. DUX4 and DUX4 target mRNA levels are typical of FSHD cells when siDUX4 is substituted with siCTRL. (N.D., not detected). (E) Immunostaining of endogenous DUX4 target ZNF217 (red) co-localizes with BFP fluorescence (blue) during FSHD myoblast differentiation.
    Figure Legend Snippet: Characterization of a DUX4-activated fluorescent reporter. ( A ) A schematic diagram of the lentiviral vector encoding a DUX4 reporter with five unique DUX4 binding site sequences, identified from individual DUX4 genomic targets and a TATA box located upstream of the sequence for nuclear-localized Blue Fluorescent Protein (nuBFP). A pTK and neomycin phosphotransferase gene (NEO) were included to allow for selection of transduced cells. ( B ) BFP is specifically present in MHC-positive myotubes (green) derived from individuals with FSHD and co-localizes with DUX4 protein (red) by immunofluorescence. ( C ) Delivery of an siRNA targeting the DUX4 transcript (siDUX4) during FSHD myoblast differentiation prohibits BFP reporter activation by eliminating DUX4 protein expression. A universal non-targeting control siRNA (siCTRL) transfected in parallel results in BFP and DUX4 expression typical of FSHD myotubes. ( D ) qRT-PCR of DUX4 and DUX4 targets BFP, CCNA1 and MBD3L2 shows reduced mRNA levels after siDUX4 delivery during FSHD myoblast differentiation. DUX4 and DUX4 target mRNA levels are typical of FSHD cells when siDUX4 is substituted with siCTRL. (N.D., not detected). (E) Immunostaining of endogenous DUX4 target ZNF217 (red) co-localizes with BFP fluorescence (blue) during FSHD myoblast differentiation.

    Techniques Used: Plasmid Preparation, Binding Assay, Sequencing, Selection, Derivative Assay, Immunofluorescence, Activation Assay, Expressing, Transfection, Quantitative RT-PCR, Immunostaining, Fluorescence

    22) Product Images from "ASBEL, an ANA/BTG3 antisense transcript required for tumorigenicity of ovarian carcinoma"

    Article Title: ASBEL, an ANA/BTG3 antisense transcript required for tumorigenicity of ovarian carcinoma

    Journal: Scientific Reports

    doi: 10.1038/srep01305

    Knockdown of ASBEL induces apoptosis of ovarian cancer cell lines. (a) qRT-PCR analysis of ASBEL expression in human ovarian cancerous and corresponding non-cancerous tissues. 1~5, serous adenocarcinoma; 6 and 7, endometrioid adenocarcinoma; 8 and 9, clear cell adenocarcinoma; 10, mucinous adenocarcinoma ; 11, dysgerminoma. Prior to fold-change calculation, the values were normalized to the signal generated from GAPDH mRNA. (b) qRT-PCR analysis of ASBEL expression in JHOC5 cells infected with a lentivirus harbouring an shRNA targeting ASBEL . Error bars represent the s.d. (n = 3). (c) Viability of ovarian cancer cell lines infected with a lentivirus expressing an shRNA targeting ASBEL was assessed by MTT assays. *, P
    Figure Legend Snippet: Knockdown of ASBEL induces apoptosis of ovarian cancer cell lines. (a) qRT-PCR analysis of ASBEL expression in human ovarian cancerous and corresponding non-cancerous tissues. 1~5, serous adenocarcinoma; 6 and 7, endometrioid adenocarcinoma; 8 and 9, clear cell adenocarcinoma; 10, mucinous adenocarcinoma ; 11, dysgerminoma. Prior to fold-change calculation, the values were normalized to the signal generated from GAPDH mRNA. (b) qRT-PCR analysis of ASBEL expression in JHOC5 cells infected with a lentivirus harbouring an shRNA targeting ASBEL . Error bars represent the s.d. (n = 3). (c) Viability of ovarian cancer cell lines infected with a lentivirus expressing an shRNA targeting ASBEL was assessed by MTT assays. *, P

    Techniques Used: Quantitative RT-PCR, Expressing, Generated, Infection, shRNA, MTT Assay

    ASBEL downregulates ANA/BTG3 protein, but not mRNA expression. (a) qRT-PCR analysis of ASBEL and ANA/BTG3 expression in ovarian cancer cell lines transfected with siRNA targeting ASBEL . Prior to fold-change calculation, the values were normalized to the signal generated from GAPDH mRNA. (b) Cell lysates from ovarian cancer cell lines transfected with siRNA targeting ASBEL were subjected to immunoblotting analysis with antibodies against the indicated proteins. α-tubulin was used as a loading control. (c) (Left) qRT-PCR analysis of ANA/BTG3 expression in JHOC5 cells transfected with either the sense or antisense ASBEL expression plasmid. (Right) Cell lysates were subjected to immunoblotting analysis with antibodies against the indicated proteins. α-tubulin was used as a loading control. (d) qRT-PCR analysis of ASBEL and ANA/BTG3 expression in JHOC5 cells infected with a lentivirus expressing an shRNA targeting ANA/BTG3. (e) JHOC5 cells that had been infected with a lentivirus expressing shRNA targeting ANA/BTG3 was transfected with siRNA targeting ASBEL and their viability was assessed. *, P
    Figure Legend Snippet: ASBEL downregulates ANA/BTG3 protein, but not mRNA expression. (a) qRT-PCR analysis of ASBEL and ANA/BTG3 expression in ovarian cancer cell lines transfected with siRNA targeting ASBEL . Prior to fold-change calculation, the values were normalized to the signal generated from GAPDH mRNA. (b) Cell lysates from ovarian cancer cell lines transfected with siRNA targeting ASBEL were subjected to immunoblotting analysis with antibodies against the indicated proteins. α-tubulin was used as a loading control. (c) (Left) qRT-PCR analysis of ANA/BTG3 expression in JHOC5 cells transfected with either the sense or antisense ASBEL expression plasmid. (Right) Cell lysates were subjected to immunoblotting analysis with antibodies against the indicated proteins. α-tubulin was used as a loading control. (d) qRT-PCR analysis of ASBEL and ANA/BTG3 expression in JHOC5 cells infected with a lentivirus expressing an shRNA targeting ANA/BTG3. (e) JHOC5 cells that had been infected with a lentivirus expressing shRNA targeting ANA/BTG3 was transfected with siRNA targeting ASBEL and their viability was assessed. *, P

    Techniques Used: Expressing, Quantitative RT-PCR, Transfection, Generated, Plasmid Preparation, Infection, shRNA

    23) Product Images from "Comparative analysis of primary metabolites and transcriptome changes between ungrafted and pumpkin-grafted watermelon during fruit development"

    Article Title: Comparative analysis of primary metabolites and transcriptome changes between ungrafted and pumpkin-grafted watermelon during fruit development

    Journal: PeerJ

    doi: 10.7717/peerj.8259

    Correlation analysis of qRT-PCR and RNA-Seq data.
    Figure Legend Snippet: Correlation analysis of qRT-PCR and RNA-Seq data.

    Techniques Used: Quantitative RT-PCR, RNA Sequencing Assay

    24) Product Images from "Transcriptional profiling of trait deterioration in the insect pathogenic nematode Heterorhabditis bacteriophora"

    Article Title: Transcriptional profiling of trait deterioration in the insect pathogenic nematode Heterorhabditis bacteriophora

    Journal: BMC Genomics

    doi: 10.1186/1471-2164-10-609

    Comparison of expression of representative genes selected from microarray data with qRT-PCR . Comparison of fold change values from microarray data with expression ratios calculated from qRT-PCR. Values were determined using qRT-PCR and represents relative expression of genes between L5M and OHB. The relative expression of the target gene ( Hb-sec-23 : Yeast sec homolog, Hb-co-II : Cytochrome c oxidase II, Hb-dao-3 : Dauer or aging adult overexpression, Hb-unc-68 : Uncoordinated, Hb-asp-3 : Aspartyl protease, Hb-hid-1 : High temperature induced dauer formation, Hb-fat-2 : Fatty acid desaturase, Hb-daf-21 : Abnormal dauer formation, Hb-rab-33 : RAB family member, Hb-spl-1 : Sphingosine-1-phosphate lyase) normalized to Hb-18s :18S rRNA and relative to the expression of control. Bars represent standard errors calculated from 4 replicates of each experiment. *Significant difference ( P
    Figure Legend Snippet: Comparison of expression of representative genes selected from microarray data with qRT-PCR . Comparison of fold change values from microarray data with expression ratios calculated from qRT-PCR. Values were determined using qRT-PCR and represents relative expression of genes between L5M and OHB. The relative expression of the target gene ( Hb-sec-23 : Yeast sec homolog, Hb-co-II : Cytochrome c oxidase II, Hb-dao-3 : Dauer or aging adult overexpression, Hb-unc-68 : Uncoordinated, Hb-asp-3 : Aspartyl protease, Hb-hid-1 : High temperature induced dauer formation, Hb-fat-2 : Fatty acid desaturase, Hb-daf-21 : Abnormal dauer formation, Hb-rab-33 : RAB family member, Hb-spl-1 : Sphingosine-1-phosphate lyase) normalized to Hb-18s :18S rRNA and relative to the expression of control. Bars represent standard errors calculated from 4 replicates of each experiment. *Significant difference ( P

    Techniques Used: Expressing, Microarray, Quantitative RT-PCR, Size-exclusion Chromatography, Over Expression

    Correlation between the fold change values from microarray and the expression ratios calculated from qRT-PCR presented as level of gene expression . Correlation coefficient between the fold change values from microarray and qRT-PCR.
    Figure Legend Snippet: Correlation between the fold change values from microarray and the expression ratios calculated from qRT-PCR presented as level of gene expression . Correlation coefficient between the fold change values from microarray and qRT-PCR.

    Techniques Used: Microarray, Expressing, Quantitative RT-PCR

    25) Product Images from "A20 plays a critical role in the immunoregulatory function of mesenchymal stem cells"

    Article Title: A20 plays a critical role in the immunoregulatory function of mesenchymal stem cells

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.12849

    Proliferation and differentiation capacities of sh CTRL C3 MSC s and shA20 C3 MSC s. ( A ) BrdU incorporation coupled with 7 AAD staining was analysed by flow cytometry. C3 MSC s were cultured in ( B ) osteogenic or ( C ) adipogenic differentiation medium for 14 days, and stained with ALP (bar = 100 μm) or Oil Red O (bar = 50 μm). ( D ) C3 MSC s cultured in osteogenic or adipogenic medium for the indicated times were analysed for ALP , Osterix, PPAR ‐γ and aP 2 by qRT ‐ PCR , ** P
    Figure Legend Snippet: Proliferation and differentiation capacities of sh CTRL C3 MSC s and shA20 C3 MSC s. ( A ) BrdU incorporation coupled with 7 AAD staining was analysed by flow cytometry. C3 MSC s were cultured in ( B ) osteogenic or ( C ) adipogenic differentiation medium for 14 days, and stained with ALP (bar = 100 μm) or Oil Red O (bar = 50 μm). ( D ) C3 MSC s cultured in osteogenic or adipogenic medium for the indicated times were analysed for ALP , Osterix, PPAR ‐γ and aP 2 by qRT ‐ PCR , ** P

    Techniques Used: BrdU Incorporation Assay, Staining, Flow Cytometry, Cytometry, Cell Culture, ALP Assay, Quantitative RT-PCR

    Morphological and phenotypic characterization of C3 MSC s after A20 knockdown. ( A ) qRT ‐ PCR analysis of A20 mRNA levels with or without A20 knockdown. ( B ) Morphology and ( C ) size of cultured sh CTRL C3 MSC s and shA20 C3 MSC s were analysed by microscopy and flow cytometry. ( D ) Cell surface markers were analysed by flow cytometry, ** P
    Figure Legend Snippet: Morphological and phenotypic characterization of C3 MSC s after A20 knockdown. ( A ) qRT ‐ PCR analysis of A20 mRNA levels with or without A20 knockdown. ( B ) Morphology and ( C ) size of cultured sh CTRL C3 MSC s and shA20 C3 MSC s were analysed by microscopy and flow cytometry. ( D ) Cell surface markers were analysed by flow cytometry, ** P

    Techniques Used: Quantitative RT-PCR, Cell Culture, Microscopy, Flow Cytometry, Cytometry

    Inflammatory cytokines induce A20 expression in MSC s. MSC s ( A ) and C3H/10T1/2 ( B and C ) were treated with 0, 2, 5 and 10 ng/ml IFN ‐γ and TNF ‐α for 24 hrs. A20 mRNA and protein levels were examined by qRT ‐ PCR and Western blot analysis. MSC s ( D ) and C3H/10T1/2 ( E and F ) were treated with 5 ng/ml IFN ‐γ and TNF ‐α for 0, 6, 12 and 24 hrs, and mRNA and protein expression levels were determined by qRT ‐ PCR and Western blot analysis, respectively.
    Figure Legend Snippet: Inflammatory cytokines induce A20 expression in MSC s. MSC s ( A ) and C3H/10T1/2 ( B and C ) were treated with 0, 2, 5 and 10 ng/ml IFN ‐γ and TNF ‐α for 24 hrs. A20 mRNA and protein levels were examined by qRT ‐ PCR and Western blot analysis. MSC s ( D ) and C3H/10T1/2 ( E and F ) were treated with 5 ng/ml IFN ‐γ and TNF ‐α for 0, 6, 12 and 24 hrs, and mRNA and protein expression levels were determined by qRT ‐ PCR and Western blot analysis, respectively.

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot

    A20 inhibits TNF ‐α and promotes IL ‐10 production in C3 MSC s, and the p38/ MAPK pathway is involved in A20‐induced immunomodulation. TNF ‐α ( A ) and IL ‐10 ( B , left) expression was examined with or without stimulation with 5 ng/ml IFN ‐γ and TNF ‐α by qRT ‐ PCR . IL ‐10 protein level was determined by ELISA ( B , right). ( C ) The time courses of p38 phosphorylation in response to 5 ng/ml IFN ‐γ and TNF ‐α was determined by immunoblotting, ** P
    Figure Legend Snippet: A20 inhibits TNF ‐α and promotes IL ‐10 production in C3 MSC s, and the p38/ MAPK pathway is involved in A20‐induced immunomodulation. TNF ‐α ( A ) and IL ‐10 ( B , left) expression was examined with or without stimulation with 5 ng/ml IFN ‐γ and TNF ‐α by qRT ‐ PCR . IL ‐10 protein level was determined by ELISA ( B , right). ( C ) The time courses of p38 phosphorylation in response to 5 ng/ml IFN ‐γ and TNF ‐α was determined by immunoblotting, ** P

    Techniques Used: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    26) Product Images from "Generation of Pigs Resistant to Highly Pathogenic-Porcine Reproductive and Respiratory Syndrome Virus through Gene Editing of CD163"

    Article Title: Generation of Pigs Resistant to Highly Pathogenic-Porcine Reproductive and Respiratory Syndrome Virus through Gene Editing of CD163

    Journal: International Journal of Biological Sciences

    doi: 10.7150/ijbs.25862

    CD163 Mut/Mut PAMs are remarkably resistant to HP-PRRSV infection . (A) After infection with the HP-PRRSV strain JXwn06 at the indicated MOIs (0.005, 0.025, 0.1, 0.25, 2.0), culture supernatants were collected at 36 hpi, and viral titers were analyzed by a standard TCID 50 assay (left). Cells were collected to measure relative expression of viral RNA by qRT-PCR (right). GAPDH mRNA was used as an endogenous control. (B) After infection with HP-PRRSV strain JXwn06 at an MOI of 0.1, viral titers were measured by TCID 50 at the indicated time points (12, 24, 36 and 48 h) (left). Relative expression of viral RNA was analyzed using qRT-PCR (right). GAPDH mRNA was used as an endogenous control. (C) PAMs were infected with JXwn06 at an MOI of 0.1, and 36 h later, levels of PRRSV protein GP5 were analyzed by Western blotting analysis (left). Expression of α-tubulin was shown as a loading control. After 48 h, cells were fixed for detection of PRRSV N protein (Green) by immunofluorescent staining(right). The nuclei (blue) were stained with DAPI. (D) The in vitro infection experiment was carried out with the HP-PRRSV strain WUH3. At the indicated MOIs (0.005, 0.025, 0.1, 0.25, 1.0), viral titers were analyzed by a standard TCID 50 assay (left). After infection at an MOI of 0.025, relative expression of viral RNA was analyzed using qRT-PCR at the indicated time points (12, 24, 36, 48, 60 and 72 h). GAPDH mRNA was used as an endogenous control. Data are presented as the mean±SD, n=3. * P
    Figure Legend Snippet: CD163 Mut/Mut PAMs are remarkably resistant to HP-PRRSV infection . (A) After infection with the HP-PRRSV strain JXwn06 at the indicated MOIs (0.005, 0.025, 0.1, 0.25, 2.0), culture supernatants were collected at 36 hpi, and viral titers were analyzed by a standard TCID 50 assay (left). Cells were collected to measure relative expression of viral RNA by qRT-PCR (right). GAPDH mRNA was used as an endogenous control. (B) After infection with HP-PRRSV strain JXwn06 at an MOI of 0.1, viral titers were measured by TCID 50 at the indicated time points (12, 24, 36 and 48 h) (left). Relative expression of viral RNA was analyzed using qRT-PCR (right). GAPDH mRNA was used as an endogenous control. (C) PAMs were infected with JXwn06 at an MOI of 0.1, and 36 h later, levels of PRRSV protein GP5 were analyzed by Western blotting analysis (left). Expression of α-tubulin was shown as a loading control. After 48 h, cells were fixed for detection of PRRSV N protein (Green) by immunofluorescent staining(right). The nuclei (blue) were stained with DAPI. (D) The in vitro infection experiment was carried out with the HP-PRRSV strain WUH3. At the indicated MOIs (0.005, 0.025, 0.1, 0.25, 1.0), viral titers were analyzed by a standard TCID 50 assay (left). After infection at an MOI of 0.025, relative expression of viral RNA was analyzed using qRT-PCR at the indicated time points (12, 24, 36, 48, 60 and 72 h). GAPDH mRNA was used as an endogenous control. Data are presented as the mean±SD, n=3. * P

    Techniques Used: Infection, Expressing, Quantitative RT-PCR, Western Blot, Staining, In Vitro

    27) Product Images from "Long noncoding RNA UPAT promotes colon tumorigenesis by inhibiting degradation of UHRF1"

    Article Title: Long noncoding RNA UPAT promotes colon tumorigenesis by inhibiting degradation of UHRF1

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1500992113

    UPAT is required for the tumorigenicity and the expression of RAS-, CDH1-, and hypoxia-related genes in colon cancer cells. ( A ) qRT-PCR analysis of UPAT expression in HCT116 cells infected with a lentivirus harboring an sh UPAT . Results are expressed as
    Figure Legend Snippet: UPAT is required for the tumorigenicity and the expression of RAS-, CDH1-, and hypoxia-related genes in colon cancer cells. ( A ) qRT-PCR analysis of UPAT expression in HCT116 cells infected with a lentivirus harboring an sh UPAT . Results are expressed as

    Techniques Used: Expressing, Quantitative RT-PCR, Infection

    UPAT stabilizes UHRF1 protein by interfering with its ubiquitination and degradation. ( A , Upper ) qRT-PCR analysis of UHRF1 expression in HCT116 cells transfected with siRNA targeting UPAT . Results are expressed as the mean ± SEM ( n = 3). ( A ,
    Figure Legend Snippet: UPAT stabilizes UHRF1 protein by interfering with its ubiquitination and degradation. ( A , Upper ) qRT-PCR analysis of UHRF1 expression in HCT116 cells transfected with siRNA targeting UPAT . Results are expressed as the mean ± SEM ( n = 3). ( A ,

    Techniques Used: Quantitative RT-PCR, Expressing, Transfection

    28) Product Images from "?-catenin promotes the type I IFN synthesis and the IFN-dependent signaling response but is suppressed by influenza A virus-induced RIG-I/NF-?B signaling"

    Article Title: ?-catenin promotes the type I IFN synthesis and the IFN-dependent signaling response but is suppressed by influenza A virus-induced RIG-I/NF-?B signaling

    Journal: Cell Communication and Signaling : CCS

    doi: 10.1186/1478-811X-12-29

    The ISG promoter activity is triggered by β- and γ-catenin. (A) A549 cells were transfected with β-catenin and LEF1 for 30 h, and the mRNA level of the type I and type III IFN-dependent MX1 gene was measured by qRT-PCR. The mRNA amount of empty vector-transfected cells was taken as unity. (B and C) Vero cells transfected for 24 h with indicated plasmids were infected with vesicular stomatitis virus (VSV) (MOI = 0.0001) for an additional 24 h. Subsequently, the overexpression of β-catenin was confirmed by immunoblotting of corresponding RIPA lysates (B) and the propagation of VSV by standard plaque titration assay (C) . (D and E) Vero cells were co-transfected with the ISRE luciferase reporter gene and plasmids coding for proteins indicated in column legends. After 24 hours, Vero cells were left unstimulated or treated with 100 U/ml IFN-β for 8 h. The y-axis represents the relative reporter gene activity with luciferase activity of unstimulated, empty vector-transfected cells being set to one. (F) Vero cells were transfected with the ISRE luciferase reporter gene, and its activity in β-catenin- and LEF1-overexpressing cells was measured in the presence or absence of co-transfected p300. The luciferase activity of β-catenin and LEF1-transfected cells was arbitrarily taken as unity. (G) A549 cells were transfected with empty vector or plasmids coding for β-catenin and LEF1 for 30 h, and the interaction of cellular proteins with the DNA was analyzed by ChIP assays using specific antibodies to IRF3 or β-catenin. The co-immunoprecipitated DNA was amplified by qRT-PCR using specific primers for the promoter region of the MX1 gene and is given as the n-fold amount to the IgG control. Representative values from one of three repeated experiments are depicted.
    Figure Legend Snippet: The ISG promoter activity is triggered by β- and γ-catenin. (A) A549 cells were transfected with β-catenin and LEF1 for 30 h, and the mRNA level of the type I and type III IFN-dependent MX1 gene was measured by qRT-PCR. The mRNA amount of empty vector-transfected cells was taken as unity. (B and C) Vero cells transfected for 24 h with indicated plasmids were infected with vesicular stomatitis virus (VSV) (MOI = 0.0001) for an additional 24 h. Subsequently, the overexpression of β-catenin was confirmed by immunoblotting of corresponding RIPA lysates (B) and the propagation of VSV by standard plaque titration assay (C) . (D and E) Vero cells were co-transfected with the ISRE luciferase reporter gene and plasmids coding for proteins indicated in column legends. After 24 hours, Vero cells were left unstimulated or treated with 100 U/ml IFN-β for 8 h. The y-axis represents the relative reporter gene activity with luciferase activity of unstimulated, empty vector-transfected cells being set to one. (F) Vero cells were transfected with the ISRE luciferase reporter gene, and its activity in β-catenin- and LEF1-overexpressing cells was measured in the presence or absence of co-transfected p300. The luciferase activity of β-catenin and LEF1-transfected cells was arbitrarily taken as unity. (G) A549 cells were transfected with empty vector or plasmids coding for β-catenin and LEF1 for 30 h, and the interaction of cellular proteins with the DNA was analyzed by ChIP assays using specific antibodies to IRF3 or β-catenin. The co-immunoprecipitated DNA was amplified by qRT-PCR using specific primers for the promoter region of the MX1 gene and is given as the n-fold amount to the IgG control. Representative values from one of three repeated experiments are depicted.

    Techniques Used: Activity Assay, Transfection, Quantitative RT-PCR, Plasmid Preparation, Infection, Over Expression, Titration, Luciferase, Chromatin Immunoprecipitation, Immunoprecipitation, Amplification

    β-catenin and LEF1 regulate IFN-β promoter activity via IRF-PRD. (A) Vero cells were transfected with the reporter gene plasmid containing the IRF3-responsive elements of the IFN-β enhanceosome along with indicated plasmids, and the reporter gene activity was measured 30 h later. The luciferase activity of β-catenin and LEF1-transfected cells was arbitrarily taken as unity. (B) A549 cells were transfected with empty vector or plasmids coding for β-catenin and LEF1 for 30 h, and the interaction of cellular proteins with the DNA was analyzed by ChIP assays. The amount of amplified DNA in IRF3- and β-catenin-specific immunoprecipitates was quantified by qRT-PCR using primers specific for the promoter region of the IFNB1 gene. Values represent n-folds of IgG controls. One of three independently repeated experiments is depicted as representative.
    Figure Legend Snippet: β-catenin and LEF1 regulate IFN-β promoter activity via IRF-PRD. (A) Vero cells were transfected with the reporter gene plasmid containing the IRF3-responsive elements of the IFN-β enhanceosome along with indicated plasmids, and the reporter gene activity was measured 30 h later. The luciferase activity of β-catenin and LEF1-transfected cells was arbitrarily taken as unity. (B) A549 cells were transfected with empty vector or plasmids coding for β-catenin and LEF1 for 30 h, and the interaction of cellular proteins with the DNA was analyzed by ChIP assays. The amount of amplified DNA in IRF3- and β-catenin-specific immunoprecipitates was quantified by qRT-PCR using primers specific for the promoter region of the IFNB1 gene. Values represent n-folds of IgG controls. One of three independently repeated experiments is depicted as representative.

    Techniques Used: Activity Assay, Transfection, Plasmid Preparation, Luciferase, Chromatin Immunoprecipitation, Amplification, Quantitative RT-PCR

    29) Product Images from "Insight into the Regulatory Relationships between the Insulin-Like Androgenic Gland Hormone Gene and the Insulin-Like Androgenic Gland Hormone-binding Protein Gene in Giant Freshwater Prawns (Macrobrachium rosenbergii)"

    Article Title: Insight into the Regulatory Relationships between the Insulin-Like Androgenic Gland Hormone Gene and the Insulin-Like Androgenic Gland Hormone-binding Protein Gene in Giant Freshwater Prawns (Macrobrachium rosenbergii)

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21124207

    Effects of dsRNA- Mr - IAG injection. ( A ) Effect of Mr - IAG dsRNA on Mr - IAG mRNA levels in the androgenic gland. ( B ) Effects of Mr - IAG dsRNA on Mr - IAGBP mRNA levels revealed by qRT-PCR in different tissues. Mr - IAG and Mr - IAGBP mRNA levels were normalized to β-actin. The tissues included: heart (Ht), testis (Te), eyestalk (Es), nerve cord (Nc), muscle (Mu), androgenic gland (Ag), hepatopancreas (Hp), and brain (Br). qRT-PCR data are shown as means ± SE (standard error). ∗ p
    Figure Legend Snippet: Effects of dsRNA- Mr - IAG injection. ( A ) Effect of Mr - IAG dsRNA on Mr - IAG mRNA levels in the androgenic gland. ( B ) Effects of Mr - IAG dsRNA on Mr - IAGBP mRNA levels revealed by qRT-PCR in different tissues. Mr - IAG and Mr - IAGBP mRNA levels were normalized to β-actin. The tissues included: heart (Ht), testis (Te), eyestalk (Es), nerve cord (Nc), muscle (Mu), androgenic gland (Ag), hepatopancreas (Hp), and brain (Br). qRT-PCR data are shown as means ± SE (standard error). ∗ p

    Techniques Used: Injection, Quantitative RT-PCR

    Transcriptional levels of Mr-IAG ( A ) and Mr-IAGBP ( B ) at different developmental stages, revealed by qRT-PCR. Mr-IAG and Mr-IAGBP mRNA levels were normalized to β-actin. qRT-PCR data are shown as means ± SE (standard error). CS, cleavage stage; BS, blastula stage; GS, gastrula stage; NS, nauplius stage; ZS, zoea stage; LH1, first-day larva after hatching; LH10, fifth-day larva after hatching; PL1, first-day post-larval stage; PL10, 10th-day post-larval stage; PL20, 20th-day post-larval stage; PL30, 30th-day post-larval stage; PL50, 50th-day post-larval stage; AD, adult stage.
    Figure Legend Snippet: Transcriptional levels of Mr-IAG ( A ) and Mr-IAGBP ( B ) at different developmental stages, revealed by qRT-PCR. Mr-IAG and Mr-IAGBP mRNA levels were normalized to β-actin. qRT-PCR data are shown as means ± SE (standard error). CS, cleavage stage; BS, blastula stage; GS, gastrula stage; NS, nauplius stage; ZS, zoea stage; LH1, first-day larva after hatching; LH10, fifth-day larva after hatching; PL1, first-day post-larval stage; PL10, 10th-day post-larval stage; PL20, 20th-day post-larval stage; PL30, 30th-day post-larval stage; PL50, 50th-day post-larval stage; AD, adult stage.

    Techniques Used: Quantitative RT-PCR

    Transcriptional levels of Mr-IAG ( A ) and Mr-IAGBP ( B ) revealed by qRT-PCR in different tissues. The tissues included: heart (Ht), testis (Te), eyestalk (Es), nerve cord (Nc), muscle (Mu), androgenic gland (Ag), hepatopancreas (Hp), and brain (Br). Mr-IAG and Mr-IAGBP mRNA levels were normalized to β-actin.
    Figure Legend Snippet: Transcriptional levels of Mr-IAG ( A ) and Mr-IAGBP ( B ) revealed by qRT-PCR in different tissues. The tissues included: heart (Ht), testis (Te), eyestalk (Es), nerve cord (Nc), muscle (Mu), androgenic gland (Ag), hepatopancreas (Hp), and brain (Br). Mr-IAG and Mr-IAGBP mRNA levels were normalized to β-actin.

    Techniques Used: Quantitative RT-PCR

    Effects of dsRNA- Mr - IAGBP injection. ( A ) Effect of Mr - IAGBP dsRNA on Mr - IAG mRNA levels in the androgenic gland. ( B ) Effect of Mr - IAGBP dsRNA on Mr - IAGBP mRNA levels in the androgenic gland. mRNA levels were analyzed by qRT-PCR. Mr - IAGBP and Mr - IAG mRNA levels were normalized to β-actin. qRT-PCR data are shown as means ± SE (standard error). ∗∗ p
    Figure Legend Snippet: Effects of dsRNA- Mr - IAGBP injection. ( A ) Effect of Mr - IAGBP dsRNA on Mr - IAG mRNA levels in the androgenic gland. ( B ) Effect of Mr - IAGBP dsRNA on Mr - IAGBP mRNA levels in the androgenic gland. mRNA levels were analyzed by qRT-PCR. Mr - IAGBP and Mr - IAG mRNA levels were normalized to β-actin. qRT-PCR data are shown as means ± SE (standard error). ∗∗ p

    Techniques Used: Injection, Quantitative RT-PCR

    30) Product Images from "Genomic Identification, Evolution, and Expression Analysis of Collagen Genes Family in Water Buffalo during Lactation"

    Article Title: Genomic Identification, Evolution, and Expression Analysis of Collagen Genes Family in Water Buffalo during Lactation

    Journal: Genes

    doi: 10.3390/genes11050515

    Heat map ( A ) of orthologous collagen genes of cattle and buffalo and validation of selected buffalo collagens by qRT-PCR ( B ). Distances, representing the relative similarity among genes and tissues, were calculated using Pearson’s correlation coefficients. Color represents the TPM (transcripts per million) values of gene expression after scaling and centering. D7 indicates early lactation, D140 indicates mid-lactation, and D280 indicates late lactation.
    Figure Legend Snippet: Heat map ( A ) of orthologous collagen genes of cattle and buffalo and validation of selected buffalo collagens by qRT-PCR ( B ). Distances, representing the relative similarity among genes and tissues, were calculated using Pearson’s correlation coefficients. Color represents the TPM (transcripts per million) values of gene expression after scaling and centering. D7 indicates early lactation, D140 indicates mid-lactation, and D280 indicates late lactation.

    Techniques Used: Quantitative RT-PCR, Expressing

    31) Product Images from "Chemosensitization of prostate cancer stem cells in mice by angiogenin and plexin-B2 inhibitors"

    Article Title: Chemosensitization of prostate cancer stem cells in mice by angiogenin and plexin-B2 inhibitors

    Journal: Communications Biology

    doi: 10.1038/s42003-020-0750-6

    Effect of ANG and PLXNB2 mAbs on chemo-sensitivity of CSC tumors. a PLXNB2 mAb sensitizes CSC tumors to DTX. NSG mice were inoculated with 100,000 CSCs, randomized into 6 groups ( n = 6–19) when tumor sizes reached ~100 mm 3 , and treated with PBS control ( n = 6), 4.8 mg/kg mAb17 ( n = 19), 10 mg/kg DTX ( n = 16), 30 mg/kg DTX ( n = 6), 10 mg/kg DTX + 4.8 mg/kg mAb17 ( n = 16), and 30 mg/kg DTX + 4.8 mg/kg mAb17 ( n = 10), by weekly i.p. injection. Tumor sizes were measured weekly. b Treatment was ceased on week 4. Animals bearing similar tumor sizes in the group of 30 mg/kg DTX (4 mice with an average tumor size of 34.4 ± 5.6 mm 3 ) and in the group of 30 mg/kg DTX + 4.8 mg/kg mAb17 (6 mice with an average tumor size of 22.2 ± 8.3 mm 3 ) were observed for tumor growth for another 23 days. c mRNA levels of ANG , PLXNB2 , and representative CSC markers in recurrent tumors determined by qRT-PCR ( n = 3). Values were normalized to the average of the DTX group. d IHC staining of CK5, CK18, and SYP in CSC xenograft tumor tissues treated with PBS or mAb17 ( n = 2). Scale bar: 50 μm. e IHC of cleaved PARP and cleaved caspase 6 in CSC xenograft tumor tissues treated with PBS, 30 mg/kg DTX, and 30 mg/kg DTX + 4.8 mg/kg mAb17 ( n = 3). Scale bar: 50 μm. f Apoptotic cells detected by TUNEL assay in CSC xenograft tumor tissues treated with DTX (30 mg/kg), mAb17 (4.8 mg/kg), or both. Scale bar: 100 μm. TUNEL positive cells were counted in a total of 200 cells in each sample ( n = 2). * p
    Figure Legend Snippet: Effect of ANG and PLXNB2 mAbs on chemo-sensitivity of CSC tumors. a PLXNB2 mAb sensitizes CSC tumors to DTX. NSG mice were inoculated with 100,000 CSCs, randomized into 6 groups ( n = 6–19) when tumor sizes reached ~100 mm 3 , and treated with PBS control ( n = 6), 4.8 mg/kg mAb17 ( n = 19), 10 mg/kg DTX ( n = 16), 30 mg/kg DTX ( n = 6), 10 mg/kg DTX + 4.8 mg/kg mAb17 ( n = 16), and 30 mg/kg DTX + 4.8 mg/kg mAb17 ( n = 10), by weekly i.p. injection. Tumor sizes were measured weekly. b Treatment was ceased on week 4. Animals bearing similar tumor sizes in the group of 30 mg/kg DTX (4 mice with an average tumor size of 34.4 ± 5.6 mm 3 ) and in the group of 30 mg/kg DTX + 4.8 mg/kg mAb17 (6 mice with an average tumor size of 22.2 ± 8.3 mm 3 ) were observed for tumor growth for another 23 days. c mRNA levels of ANG , PLXNB2 , and representative CSC markers in recurrent tumors determined by qRT-PCR ( n = 3). Values were normalized to the average of the DTX group. d IHC staining of CK5, CK18, and SYP in CSC xenograft tumor tissues treated with PBS or mAb17 ( n = 2). Scale bar: 50 μm. e IHC of cleaved PARP and cleaved caspase 6 in CSC xenograft tumor tissues treated with PBS, 30 mg/kg DTX, and 30 mg/kg DTX + 4.8 mg/kg mAb17 ( n = 3). Scale bar: 50 μm. f Apoptotic cells detected by TUNEL assay in CSC xenograft tumor tissues treated with DTX (30 mg/kg), mAb17 (4.8 mg/kg), or both. Scale bar: 100 μm. TUNEL positive cells were counted in a total of 200 cells in each sample ( n = 2). * p

    Techniques Used: Mouse Assay, Injection, Quantitative RT-PCR, Immunohistochemistry, Staining, TUNEL Assay

    Differentiation potential of prostate CSCs. a Expression levels of basal, luminal, and neuroendocrine markers in CSCs and parent cells measured by qRT-PCR ( n = 3). Values in the CSCs were normalized to the respective parent cells. b Morphology of PC3 CSCs cultured in sphere medium in non-adherent petri dish (top) and in DMEM plus 10% FBS in cell culture dish (bottom) for 2 weeks. Scale bar, 500 μm. c mRNA levels of basal, luminal, and neuroendocrine markers in CSCs cultured in regular cell culture medium (DMEM + 10% FBS) for 3 and 8 days ( n = 3). Heatmaps represent the relative mRNA level of each marker normalized to the value at day 1. d mRNA levels of basal, luminal, and neuroendocrine markers in CSCs and CSC-derived tumors (left), and in PC3 cells and PC3 cell-derived tumors (right) ( n = 3). Values in the tumor tissues were normalized to the originating cells. IHC of CK18 ( e ) and SYP ( f ) in tumors derived from PC3 cells or CSCs. Quantitation of signal intensity was obtained by Image J ( n = 3). Scale bar: left panels, 200 μm; right panels, 50 μm. ns not significant.
    Figure Legend Snippet: Differentiation potential of prostate CSCs. a Expression levels of basal, luminal, and neuroendocrine markers in CSCs and parent cells measured by qRT-PCR ( n = 3). Values in the CSCs were normalized to the respective parent cells. b Morphology of PC3 CSCs cultured in sphere medium in non-adherent petri dish (top) and in DMEM plus 10% FBS in cell culture dish (bottom) for 2 weeks. Scale bar, 500 μm. c mRNA levels of basal, luminal, and neuroendocrine markers in CSCs cultured in regular cell culture medium (DMEM + 10% FBS) for 3 and 8 days ( n = 3). Heatmaps represent the relative mRNA level of each marker normalized to the value at day 1. d mRNA levels of basal, luminal, and neuroendocrine markers in CSCs and CSC-derived tumors (left), and in PC3 cells and PC3 cell-derived tumors (right) ( n = 3). Values in the tumor tissues were normalized to the originating cells. IHC of CK18 ( e ) and SYP ( f ) in tumors derived from PC3 cells or CSCs. Quantitation of signal intensity was obtained by Image J ( n = 3). Scale bar: left panels, 200 μm; right panels, 50 μm. ns not significant.

    Techniques Used: Expressing, Quantitative RT-PCR, Cell Culture, Marker, Derivative Assay, Immunohistochemistry, Quantitation Assay

    Knockdown of ANG or PLXNB2 decreases stemness of prostate CSCs. a mRNA levels of cancer stemness-related genes in prostate CSCs and parent cells ( n = 5). Values in the CSCs were normalized to the respective parent cells. mRNA and protein levels of ANG ( b ) and PLXNB2 ( c ) in knockdown CSCs. mRNA levels were determined by qRT-PCR and normalized to control shRNA transfectants ( n = 3). Protein levels were determined by immunoblotting. d mRNA level of cancer stemness-related genes in ANG and PLXNB2 knockdown CSCs ( n = 3). Values were normalized to the control shRNA transfectants. Cell cycle status of ANG ( e ) and PLXNB2 ( f ) knockdown CSCs, analyzed by flow cytometry after Ki-67 and 7-AAD staining ( n = 2). Sphere formation of ANG ( g ) and PLXNB2 ( h ) knockdown CSCs ( n = 3). i CSC exhaustion during serial passaging in vivo. Cells were passaged in NSG mice ( n = 5–8) for three times. In each passage, 100,000 cells were inoculated per mouse. Tumors were excised and weighed 4 weeks post inoculation in each passage. Exhaustion rate was calculated as the ratio of tumor weight from first passage to third passage. * p
    Figure Legend Snippet: Knockdown of ANG or PLXNB2 decreases stemness of prostate CSCs. a mRNA levels of cancer stemness-related genes in prostate CSCs and parent cells ( n = 5). Values in the CSCs were normalized to the respective parent cells. mRNA and protein levels of ANG ( b ) and PLXNB2 ( c ) in knockdown CSCs. mRNA levels were determined by qRT-PCR and normalized to control shRNA transfectants ( n = 3). Protein levels were determined by immunoblotting. d mRNA level of cancer stemness-related genes in ANG and PLXNB2 knockdown CSCs ( n = 3). Values were normalized to the control shRNA transfectants. Cell cycle status of ANG ( e ) and PLXNB2 ( f ) knockdown CSCs, analyzed by flow cytometry after Ki-67 and 7-AAD staining ( n = 2). Sphere formation of ANG ( g ) and PLXNB2 ( h ) knockdown CSCs ( n = 3). i CSC exhaustion during serial passaging in vivo. Cells were passaged in NSG mice ( n = 5–8) for three times. In each passage, 100,000 cells were inoculated per mouse. Tumors were excised and weighed 4 weeks post inoculation in each passage. Exhaustion rate was calculated as the ratio of tumor weight from first passage to third passage. * p

    Techniques Used: Quantitative RT-PCR, shRNA, Flow Cytometry, Cytometry, Staining, Passaging, In Vivo, Mouse Assay

    32) Product Images from "Transcriptomic analysis of Citrus clementina mandarin fruits maturation reveals a MADS-box transcription factor that might be involved in the regulation of earliness"

    Article Title: Transcriptomic analysis of Citrus clementina mandarin fruits maturation reveals a MADS-box transcription factor that might be involved in the regulation of earliness

    Journal: BMC Plant Biology

    doi: 10.1186/s12870-019-1651-z

    qRT-PCR experiments. a Results of the qRT-PCR experiments, shown as expression fold change of ARR and HER relative to CLE at 30 and 126 DPA. Original RNA from peel at 126 DPA and newly extracted from fruitlets at 30 DPA were used. The quantitative analysis confirms the RNA-Seq data for Ciclev10021357 , the SlMADS1 homolog, Ciclev10032572 , Ciclev10021100, Ciclev10020575 and Ciclev10019920 . b Image of ARR, CLE and HER fruitlets at 30 DPA, where the differences in size can be appreciated, the rule on the left side shows mm. Error bars represent SEM values
    Figure Legend Snippet: qRT-PCR experiments. a Results of the qRT-PCR experiments, shown as expression fold change of ARR and HER relative to CLE at 30 and 126 DPA. Original RNA from peel at 126 DPA and newly extracted from fruitlets at 30 DPA were used. The quantitative analysis confirms the RNA-Seq data for Ciclev10021357 , the SlMADS1 homolog, Ciclev10032572 , Ciclev10021100, Ciclev10020575 and Ciclev10019920 . b Image of ARR, CLE and HER fruitlets at 30 DPA, where the differences in size can be appreciated, the rule on the left side shows mm. Error bars represent SEM values

    Techniques Used: Quantitative RT-PCR, Expressing, RNA Sequencing Assay

    33) Product Images from "Genome-wide identification and characterization of the bHLH gene family in tomato"

    Article Title: Genome-wide identification and characterization of the bHLH gene family in tomato

    Journal: BMC Genomics

    doi: 10.1186/s12864-014-1209-2

    Expression analyses of the six SlbHLH genes under iron-deficient stress by qRT-PCR. For qRT-PCR, the relative amount of mRNA (y-axis) was calculated by according to the description in Methods. The x-axis indicates the shoot (S) and root (R) of tomato under iron-sufficient (+Fe) and iron deficient (-Fe) conditions.
    Figure Legend Snippet: Expression analyses of the six SlbHLH genes under iron-deficient stress by qRT-PCR. For qRT-PCR, the relative amount of mRNA (y-axis) was calculated by according to the description in Methods. The x-axis indicates the shoot (S) and root (R) of tomato under iron-sufficient (+Fe) and iron deficient (-Fe) conditions.

    Techniques Used: Expressing, Quantitative RT-PCR

    34) Product Images from "MIR205HG acts as a ceRNA to expedite cell proliferation and progression in lung squamous cell carcinoma via targeting miR-299-3p/MAP3K2 axis"

    Article Title: MIR205HG acts as a ceRNA to expedite cell proliferation and progression in lung squamous cell carcinoma via targeting miR-299-3p/MAP3K2 axis

    Journal: BMC Pulmonary Medicine

    doi: 10.1186/s12890-020-1174-2

    MIR205HG was an upstream factor for miR-299-3p in LUSC. a . The potential miRNA expression in LUSC was downloaded from dbDEMC 2 website. b . RIP assay was applied to confirm the relationship between MIR205HG and miR-299-3p. c . StarBase database was used to predict the potential binding sites between MIR205HG and miR-299-3p. d . Luciferase reporter assay was carried out to verify that MIR205HG could bind with miR-299-3p. e . qRT-PCR analysis was performed to assess the relative expression of miR-299-3p in both cell lines transfected with sh-MIR205HG#1. All data were showed as the mean ± SD. ** P
    Figure Legend Snippet: MIR205HG was an upstream factor for miR-299-3p in LUSC. a . The potential miRNA expression in LUSC was downloaded from dbDEMC 2 website. b . RIP assay was applied to confirm the relationship between MIR205HG and miR-299-3p. c . StarBase database was used to predict the potential binding sites between MIR205HG and miR-299-3p. d . Luciferase reporter assay was carried out to verify that MIR205HG could bind with miR-299-3p. e . qRT-PCR analysis was performed to assess the relative expression of miR-299-3p in both cell lines transfected with sh-MIR205HG#1. All data were showed as the mean ± SD. ** P

    Techniques Used: Expressing, Binding Assay, Luciferase, Reporter Assay, Quantitative RT-PCR, Transfection

    Cutting down MIR205HG expression impaired cell proliferation and migration abilities in LUSC. a . GEPIA dataset was applied to examine the relative expression of MIR205HG in LUSC tissues and adjacent normal tissues. b . Carrying out qRT-PCR to determine different expressions of MIR205H in normal lung squamous cell line (BEAS-2B) and LUSC cell lines (NCI-H520, SK-MES-1 and NCI-H1703). c . The relative expression of MIR205H under the condition of down-regulating MIR205H was measured by qRT-PCR analysis. d . Number of colonies was assessed by colony formation assay to detect cell proliferation ability. e . Cell apoptosis rate was estimated by flow cytometry analysis. f . Transwell assay was used to measure cell migration capability by counting number of migrated cells. g . The expressions of the epithelial marker (E-cadherin) and the mesenchymal markers (N-cadherin and Vimentin) were measured by western blot analysis. All data were displayed as the mean ± SD. * P
    Figure Legend Snippet: Cutting down MIR205HG expression impaired cell proliferation and migration abilities in LUSC. a . GEPIA dataset was applied to examine the relative expression of MIR205HG in LUSC tissues and adjacent normal tissues. b . Carrying out qRT-PCR to determine different expressions of MIR205H in normal lung squamous cell line (BEAS-2B) and LUSC cell lines (NCI-H520, SK-MES-1 and NCI-H1703). c . The relative expression of MIR205H under the condition of down-regulating MIR205H was measured by qRT-PCR analysis. d . Number of colonies was assessed by colony formation assay to detect cell proliferation ability. e . Cell apoptosis rate was estimated by flow cytometry analysis. f . Transwell assay was used to measure cell migration capability by counting number of migrated cells. g . The expressions of the epithelial marker (E-cadherin) and the mesenchymal markers (N-cadherin and Vimentin) were measured by western blot analysis. All data were displayed as the mean ± SD. * P

    Techniques Used: Expressing, Migration, Quantitative RT-PCR, Colony Assay, Flow Cytometry, Transwell Assay, Marker, Western Blot

    35) Product Images from "Characterization of a novel compound that promotes myogenesis via Akt and transcriptional co-activator with PDZ-binding motif (TAZ) in mouse C2C12 cells"

    Article Title: Characterization of a novel compound that promotes myogenesis via Akt and transcriptional co-activator with PDZ-binding motif (TAZ) in mouse C2C12 cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0231265

    The effect of IBS004735 on the expression of myogenic marker genes. (A) qRT-PCR analysis of indicated genes. After C2C12 cells were transferred from growth medium to differentiation medium, mRNAs were collected at the indicated time points and qRT-PCRs were performed. IBS004735 enhanced the expression of myogenin ( Myog ) and MyoD ( Myod1 ) at 24 h. Connective tissue growth factor ( Ctgf ) expression was enhanced at 72 h by IBS004735. Ccnd1 expression was not affected. Data are means and standard errors of the means. ns, not significant; **, p
    Figure Legend Snippet: The effect of IBS004735 on the expression of myogenic marker genes. (A) qRT-PCR analysis of indicated genes. After C2C12 cells were transferred from growth medium to differentiation medium, mRNAs were collected at the indicated time points and qRT-PCRs were performed. IBS004735 enhanced the expression of myogenin ( Myog ) and MyoD ( Myod1 ) at 24 h. Connective tissue growth factor ( Ctgf ) expression was enhanced at 72 h by IBS004735. Ccnd1 expression was not affected. Data are means and standard errors of the means. ns, not significant; **, p

    Techniques Used: Expressing, Marker, Quantitative RT-PCR

    36) Product Images from "Sequentially induced motor neurons from human fibroblasts facilitate locomotor recovery in a rodent spinal cord injury model"

    Article Title: Sequentially induced motor neurons from human fibroblasts facilitate locomotor recovery in a rodent spinal cord injury model

    Journal: eLife

    doi: 10.7554/eLife.52069

    Primary induction of POU5F1(OCT4) outperforms simultaneous induction of POU5F1(OCT4) and LHX3 . ( A ) Representative images of the cells after POU5F1(OCT4) and POU5F1(OCT4) / LHX3 transduction, respectively. The morphological changes of the cells are shown at day 3, 7, 14, after picking, and at passage 1 (P1). Scale bars, 125 µm. ( B ) Proliferation rate of iMNICs induced by POU5F1(OCT4) and POU5F1(OCT4) / LHX3 transduction. Each point refers to the number of cells every 24 hr. Data are presented as mean ± SD, and represent experimental replicates (n = 4). ( C ) qRT-PCR analysis of mRNA expression level for ISL1 in iMNICs induced by POU5F1(OCT4) and POU5F1(OCT4) / LHX3 transduction. Graphs are normalized by GAPDH . Data are presented as mean ± SD, and represent experimental replicates (n = 3). ( D ) The number of colonies emerged after transduction of POU5F1(OCT4) or POU5F1(OCT4) / LHX3 were quantified. Data are presented as mean ± SD, and represent experimental replicates (n = 4).
    Figure Legend Snippet: Primary induction of POU5F1(OCT4) outperforms simultaneous induction of POU5F1(OCT4) and LHX3 . ( A ) Representative images of the cells after POU5F1(OCT4) and POU5F1(OCT4) / LHX3 transduction, respectively. The morphological changes of the cells are shown at day 3, 7, 14, after picking, and at passage 1 (P1). Scale bars, 125 µm. ( B ) Proliferation rate of iMNICs induced by POU5F1(OCT4) and POU5F1(OCT4) / LHX3 transduction. Each point refers to the number of cells every 24 hr. Data are presented as mean ± SD, and represent experimental replicates (n = 4). ( C ) qRT-PCR analysis of mRNA expression level for ISL1 in iMNICs induced by POU5F1(OCT4) and POU5F1(OCT4) / LHX3 transduction. Graphs are normalized by GAPDH . Data are presented as mean ± SD, and represent experimental replicates (n = 3). ( D ) The number of colonies emerged after transduction of POU5F1(OCT4) or POU5F1(OCT4) / LHX3 were quantified. Data are presented as mean ± SD, and represent experimental replicates (n = 4).

    Techniques Used: Transduction, Quantitative RT-PCR, Expressing

    Generation of iMNs from additional human fibroblasts (HF2). ( A ) The morphological changes of additional human fibroblast line (HF2) into iMNs (HF2-iMNs) during MN direct conversion process. Scale bars, 125 μm. ( B ) Immunofluorescence images of HB9:GFP+ HF2-iMNs (HF2-iMN6) co-expressing MN specific markers (ISLT1, HB9, MAP2, and CHAT). Scale bars, 125 μm. ( C ) Immunofluorescence images of HF2-iMN6 stained with TUJ1 and synaptic markers, SYN1 and SV2. Scale bars, 125 μm. ( D ) Immunofluorescence images of HB9:GFP+ HF2-iMN6 labeled with α-bungarotoxin-alexa555, showing formation of NMJs with myotubes. Scale bars, 75 μm. ( E ) Morphologies of established HF2-iMNIC clones (HF2-iMNIC1, 2, 4 and 8). Scale bars, 125 μm. ( F ) qRT-PCR analysis of ISL1 gene expression in HF2-iMNIC clones relative to fibroblasts. Graphs present fold changes after normalization to GAPDH . Data are presented as mean ± SD, and represent experimental replicates (n = 3). ( G ) The emergence of HB9:GFP+ iMNs (HF2-iMN1, 2, 4 and 8) converted from HF2-iMNIC clones. The morphologies of the cells are shown in phase contrast. Scale bars, 125 μm.
    Figure Legend Snippet: Generation of iMNs from additional human fibroblasts (HF2). ( A ) The morphological changes of additional human fibroblast line (HF2) into iMNs (HF2-iMNs) during MN direct conversion process. Scale bars, 125 μm. ( B ) Immunofluorescence images of HB9:GFP+ HF2-iMNs (HF2-iMN6) co-expressing MN specific markers (ISLT1, HB9, MAP2, and CHAT). Scale bars, 125 μm. ( C ) Immunofluorescence images of HF2-iMN6 stained with TUJ1 and synaptic markers, SYN1 and SV2. Scale bars, 125 μm. ( D ) Immunofluorescence images of HB9:GFP+ HF2-iMN6 labeled with α-bungarotoxin-alexa555, showing formation of NMJs with myotubes. Scale bars, 75 μm. ( E ) Morphologies of established HF2-iMNIC clones (HF2-iMNIC1, 2, 4 and 8). Scale bars, 125 μm. ( F ) qRT-PCR analysis of ISL1 gene expression in HF2-iMNIC clones relative to fibroblasts. Graphs present fold changes after normalization to GAPDH . Data are presented as mean ± SD, and represent experimental replicates (n = 3). ( G ) The emergence of HB9:GFP+ iMNs (HF2-iMN1, 2, 4 and 8) converted from HF2-iMNIC clones. The morphologies of the cells are shown in phase contrast. Scale bars, 125 μm.

    Techniques Used: Immunofluorescence, Expressing, Staining, Labeling, Clone Assay, Quantitative RT-PCR

    Establishment of iMNIC clones and upregulation of ISL1 expression after POU5F1(OCT4) induction. ( A ) The morphology of established iMNIC clones (HF1-iMNIC2, 5, 6, 7, 11 and 12). Scale bars, 125 μm. ( B ) qRT-PCR analysis of mRNA expression level for motor neuron genes ( ISL1 , HB9 , NKX6.1 , and LHX3 ) in cells at day 7 and 10 after POU5F1(OCT4) transduction and iMNIC clones relative to fibroblasts. Graphs present fold changes after normalization to GAPDH . Data are presented as mean ± SD, and represent experimental replicates (n = 3). ( C ) Immunofluorescence images of iMNICs stained with motor neuron markers (ISL1, HB9, NKX6.1 and LHX3), and neuronal marker (TUJ1 and MAP2). The nuclei were counterstained with DAPI. Scale bars, 125 µm.
    Figure Legend Snippet: Establishment of iMNIC clones and upregulation of ISL1 expression after POU5F1(OCT4) induction. ( A ) The morphology of established iMNIC clones (HF1-iMNIC2, 5, 6, 7, 11 and 12). Scale bars, 125 μm. ( B ) qRT-PCR analysis of mRNA expression level for motor neuron genes ( ISL1 , HB9 , NKX6.1 , and LHX3 ) in cells at day 7 and 10 after POU5F1(OCT4) transduction and iMNIC clones relative to fibroblasts. Graphs present fold changes after normalization to GAPDH . Data are presented as mean ± SD, and represent experimental replicates (n = 3). ( C ) Immunofluorescence images of iMNICs stained with motor neuron markers (ISL1, HB9, NKX6.1 and LHX3), and neuronal marker (TUJ1 and MAP2). The nuclei were counterstained with DAPI. Scale bars, 125 µm.

    Techniques Used: Clone Assay, Expressing, Quantitative RT-PCR, Transduction, Immunofluorescence, Staining, Marker

    37) Product Images from "Genome-wide analysis of cotton GH3 subfamily II reveals functional divergence in fiber development, hormone response and plant architecture"

    Article Title: Genome-wide analysis of cotton GH3 subfamily II reveals functional divergence in fiber development, hormone response and plant architecture

    Journal: BMC Plant Biology

    doi: 10.1186/s12870-018-1545-5

    Expression profile of GhGH3s based on RNA-seq and qRT-PCR in various tissue/organs/stages. a Trends in GhGH3 gene expression based on publicly available RNA-seq data. Expression of individual genes are shown for 20 GhGH3s . Numbers on the x-axis indicate days post anthesis, with negative numbers implying days before anthesis. The prefix OV is for ovules and F for fiber. Green, black and red backgrounds represent low, intermediate and high expression levels, respectively. The original RPKM (reads per kilobase per million) values are normalized to 0–1 and shown in boxes. b Pattern of gene expression by qRT-PCR analysis. Expression of allele pairs or individual genes are shown. Values on the y-axis represent relative expression levels while the x-axis indicates days post anthesis. Materials from the field under normal conditions were tagged and sampled. The relative expression levels of GhGH3 genes were measured in roots (R), stems (S), leaves (L), flowers (FL), early stage developing ovules with attached fibers (1, 3, 5 DPA OF, ovules plus fibers), and different stages of detached ovules and fibers (7, 10, 15, 20 DPA O/F, ovules or fibers). GhHis 3 was used as an internal control to normalize the expression data. Error bars show standard deviation calculated from three replicates
    Figure Legend Snippet: Expression profile of GhGH3s based on RNA-seq and qRT-PCR in various tissue/organs/stages. a Trends in GhGH3 gene expression based on publicly available RNA-seq data. Expression of individual genes are shown for 20 GhGH3s . Numbers on the x-axis indicate days post anthesis, with negative numbers implying days before anthesis. The prefix OV is for ovules and F for fiber. Green, black and red backgrounds represent low, intermediate and high expression levels, respectively. The original RPKM (reads per kilobase per million) values are normalized to 0–1 and shown in boxes. b Pattern of gene expression by qRT-PCR analysis. Expression of allele pairs or individual genes are shown. Values on the y-axis represent relative expression levels while the x-axis indicates days post anthesis. Materials from the field under normal conditions were tagged and sampled. The relative expression levels of GhGH3 genes were measured in roots (R), stems (S), leaves (L), flowers (FL), early stage developing ovules with attached fibers (1, 3, 5 DPA OF, ovules plus fibers), and different stages of detached ovules and fibers (7, 10, 15, 20 DPA O/F, ovules or fibers). GhHis 3 was used as an internal control to normalize the expression data. Error bars show standard deviation calculated from three replicates

    Techniques Used: Expressing, RNA Sequencing Assay, Quantitative RT-PCR, Standard Deviation

    Organization of regulatory elements of GhGH3s and their expression in response to phytohormones. a The regulatory region of each GhGH3 gene was analyzed, the numbers indicate the sum of various cis -acting elements response to the same stimuli. b The summary of GhGH3 expression response to treatment with IAA, SA, BL and GA in roots (R) and stems (S). Each orthologous pair from At- and Dt-subgenome of G. hirsutum was designed as a single gene and tested together due to their highly identical sequences. In total, 11 analogue genes standing for 20 GhGH3s were generated and could be divided into 3 subgroups (subgroups 1–3). The expression pattern of 11 analogue genes was detected by qRT-PCR. The relative expression intensity, that is, the ratio of the highest expression level to that of the 0 h control, was allocated into six different grades ranging from suppression shown as black boxes to over 20-fold induction shown as red boxes
    Figure Legend Snippet: Organization of regulatory elements of GhGH3s and their expression in response to phytohormones. a The regulatory region of each GhGH3 gene was analyzed, the numbers indicate the sum of various cis -acting elements response to the same stimuli. b The summary of GhGH3 expression response to treatment with IAA, SA, BL and GA in roots (R) and stems (S). Each orthologous pair from At- and Dt-subgenome of G. hirsutum was designed as a single gene and tested together due to their highly identical sequences. In total, 11 analogue genes standing for 20 GhGH3s were generated and could be divided into 3 subgroups (subgroups 1–3). The expression pattern of 11 analogue genes was detected by qRT-PCR. The relative expression intensity, that is, the ratio of the highest expression level to that of the 0 h control, was allocated into six different grades ranging from suppression shown as black boxes to over 20-fold induction shown as red boxes

    Techniques Used: Expressing, Generated, Quantitative RT-PCR

    38) Product Images from "Assessment of the Effects of Bisphenol A on Dopamine Synthesis and Blood Vessels in the Goldfish Brain"

    Article Title: Assessment of the Effects of Bisphenol A on Dopamine Synthesis and Blood Vessels in the Goldfish Brain

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20246206

    DEGs in the fluid shear stress and atherosclerosis pathway and validation by qRT-PCR. ( A ) Expression levels of 6 DEGs in the fluid shear stress and atherosclerosis pathway from the transcriptome data. ( B ) Validation of DEGs in the fluid shear stress and atherosclerosis pathway by qRT-PCR ( n = 3). CG: Control group; TG: BPA treatment group; and FPKM: Fragments per kilobase of transcript per million mapped reads. The qRT-PCR results were calculated from at least three independent experiments. EF1 was used as an internal normalization control. The data are expressed as the mean ± SD (* p
    Figure Legend Snippet: DEGs in the fluid shear stress and atherosclerosis pathway and validation by qRT-PCR. ( A ) Expression levels of 6 DEGs in the fluid shear stress and atherosclerosis pathway from the transcriptome data. ( B ) Validation of DEGs in the fluid shear stress and atherosclerosis pathway by qRT-PCR ( n = 3). CG: Control group; TG: BPA treatment group; and FPKM: Fragments per kilobase of transcript per million mapped reads. The qRT-PCR results were calculated from at least three independent experiments. EF1 was used as an internal normalization control. The data are expressed as the mean ± SD (* p

    Techniques Used: Quantitative RT-PCR, Expressing

    DEGs in the dopaminergic signaling pathway and analysis of the expression of dopaminergic signaling pathway-related genes by qRT-PCR. ( A ) Expression levels of 3 DEGs in the dopaminergic signaling pathway from the transcriptome data. ( B ) Expression levels of 6 dopaminergic signaling pathway-related genes determined by qRT-PCR ( n = 3). CG: Control group; TG: BPA treatment group; and FPKM: Fragments per kilobase of transcript per million mapped reads. The qRT-PCR results were calculated from at least three independent experiments. EF1 was used as an internal normalization control. The data are expressed as the mean ± SD (* p
    Figure Legend Snippet: DEGs in the dopaminergic signaling pathway and analysis of the expression of dopaminergic signaling pathway-related genes by qRT-PCR. ( A ) Expression levels of 3 DEGs in the dopaminergic signaling pathway from the transcriptome data. ( B ) Expression levels of 6 dopaminergic signaling pathway-related genes determined by qRT-PCR ( n = 3). CG: Control group; TG: BPA treatment group; and FPKM: Fragments per kilobase of transcript per million mapped reads. The qRT-PCR results were calculated from at least three independent experiments. EF1 was used as an internal normalization control. The data are expressed as the mean ± SD (* p

    Techniques Used: Expressing, Quantitative RT-PCR

    39) Product Images from "Overexpression of cytoplasmic sphingosine 1-phosphate receptor 1 promotes cell cycle progression and migration in human esophageal squamous cell carcinoma"

    Article Title: Overexpression of cytoplasmic sphingosine 1-phosphate receptor 1 promotes cell cycle progression and migration in human esophageal squamous cell carcinoma

    Journal: International Journal of Clinical and Experimental Pathology

    doi:

    The expression and subcellular localization of S1PR1 is different in transiently transfected Eca109 cells at different time points. A. S1PR1 protein was weakly expressed in the cytoplasm of Eca109 cells. B. A significant increase of S1PR1 mRNA in the S1PR1-EGFP-transfected Eca109 cells was confirmed by qRT-PCR. C. Cellular localization pattern of S1PR1-EGFP and Control-EGFP fusion protein was analyzed by confocal microscopy. Scale bar, 10 µm.
    Figure Legend Snippet: The expression and subcellular localization of S1PR1 is different in transiently transfected Eca109 cells at different time points. A. S1PR1 protein was weakly expressed in the cytoplasm of Eca109 cells. B. A significant increase of S1PR1 mRNA in the S1PR1-EGFP-transfected Eca109 cells was confirmed by qRT-PCR. C. Cellular localization pattern of S1PR1-EGFP and Control-EGFP fusion protein was analyzed by confocal microscopy. Scale bar, 10 µm.

    Techniques Used: Expressing, Transfection, Quantitative RT-PCR, Confocal Microscopy

    40) Product Images from "Importance of PNO1 for growth and survival of urinary bladder carcinoma: Role in core‐regulatory circuitry, et al. Importance of PNO1 for growth and survival of urinary bladder carcinoma: Role in core‐regulatory circuitry"

    Article Title: Importance of PNO1 for growth and survival of urinary bladder carcinoma: Role in core‐regulatory circuitry, et al. Importance of PNO1 for growth and survival of urinary bladder carcinoma: Role in core‐regulatory circuitry

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.14835

    PNO1 knockdown affected the proliferation and survival of T24 and 5637 bladder cells. (A) qRT‐PCR showing significantly reduced mRNA expression level of PNO1 against GAPDH in PNO1 knockdown cells. (B) Western blot showing reduced PNO1 protein expression in PNO1 knockdown cells. (C) Growth curves of T24 cells and (D) 5637 cells measured by automated cell counting. (E) Growth curves of T24 cells and (F) 5637 cells measured by MTT assay. (G) Comparison of colony forming ability between control and PNO1 knockdown cells. (H) Comparison of the percentage of apoptotic cells between control and PNO1 knockdown cells. * P
    Figure Legend Snippet: PNO1 knockdown affected the proliferation and survival of T24 and 5637 bladder cells. (A) qRT‐PCR showing significantly reduced mRNA expression level of PNO1 against GAPDH in PNO1 knockdown cells. (B) Western blot showing reduced PNO1 protein expression in PNO1 knockdown cells. (C) Growth curves of T24 cells and (D) 5637 cells measured by automated cell counting. (E) Growth curves of T24 cells and (F) 5637 cells measured by MTT assay. (G) Comparison of colony forming ability between control and PNO1 knockdown cells. (H) Comparison of the percentage of apoptotic cells between control and PNO1 knockdown cells. * P

    Techniques Used: Quantitative RT-PCR, Expressing, Western Blot, Cell Counting, MTT Assay

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    Purification:

    Article Title: A Myosin-7B dependent endocytosis pathway mediates cellular entry of α-Synuclein fibrils and polycation-bearing cargos
    Article Snippet: .. qRT-PCR analysis of knockout or knockdown cells Total RNA was extracted from 3 million HEK293T cells or 0.75 million primary neurons using TriPure reagent (Roche) and purified using RNeasy MinElute Cleanup Kit (Promega) following standard protocols. .. The RNA concentration was measured by Nanodrop 2000 UV spectrophotometer and 1µg total RNA was converted to cDNA using the iScript Reverse Transcription Supermix (BioRad) system.

    SYBR Green Assay:

    Article Title: Cellular transformation by combined lineage conversion and oncogene expression
    Article Snippet: .. For qRT-PCR analysis, cDNA synthesis from two biological replicates was performed using the Transcriptor High-fidelity cDNA synthesis kit (Roche) and real-time PCR using SYBR green (Roche) with primers specific for each transcript (Table S2). ..

    Article Title: Danger-Associated Peptide Regulates Root Immune Responses and Root Growth by Affecting ROS Formation in Arabidopsis
    Article Snippet: .. The 2 μg RNA was used to synthesis the cDNA by using M-MLV Reverse Transcriptase (Promega, Madison, WI, USA). qRT-PCR analysis was performed using the FastStart Universal SYBR Green mastermix (Roche Diagnostics, Hong Kong) on a CFX Connect Real Time System (Bio-Rad, Berkeley, CA, USA) using Actin2 as internal standards. ..

    Article Title: Upregulation of erythropoietin and erythropoietin receptor in castration-resistant progression of prostate cancer
    Article Snippet: .. For qRT-PCR analysis, cDNA was amplified with a SYBR Green PCR Kit (Roche) on an ABI PRISM 7900 sequence detection system, with the following EPO primers (sense 5'-ACC AAC ATT GCT TGT GCC AC-3' and antisense 5'-TCT GAA TGC TTC CTG CTC TGG-3') and EPOR primers (sense 5'-ACC GTG TCA TCC ACA TCA AT-3' and antisense 5'-GCC TTC AAA CTC GCT CTC TG-3'). .. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was amplified as an internal control.

    Polymerase Chain Reaction:

    Article Title: A member of the ETS family, EHF, and the ATPase RUVBL1 inhibit p53-mediated apoptosis
    Article Snippet: .. Total RNA was isolated using the TRIsure (BIOLINE), and 1 mg RNA was reverse transcribed with SuperScript III Reverse transcriptase (Invitrogen) using an oligo(dT)18 primer. qRT–PCR analysis of cDNA and genomic DNA was performed on a LightCycler 480 (Roche Applied Science) with Syber Green PCR mastermix (Applied Biosystems). .. Primers for mRNA expression are shown in online.

    Article Title: Upregulation of erythropoietin and erythropoietin receptor in castration-resistant progression of prostate cancer
    Article Snippet: .. For qRT-PCR analysis, cDNA was amplified with a SYBR Green PCR Kit (Roche) on an ABI PRISM 7900 sequence detection system, with the following EPO primers (sense 5'-ACC AAC ATT GCT TGT GCC AC-3' and antisense 5'-TCT GAA TGC TTC CTG CTC TGG-3') and EPOR primers (sense 5'-ACC GTG TCA TCC ACA TCA AT-3' and antisense 5'-GCC TTC AAA CTC GCT CTC TG-3'). .. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was amplified as an internal control.

    Knock-Out:

    Article Title: A Myosin-7B dependent endocytosis pathway mediates cellular entry of α-Synuclein fibrils and polycation-bearing cargos
    Article Snippet: .. qRT-PCR analysis of knockout or knockdown cells Total RNA was extracted from 3 million HEK293T cells or 0.75 million primary neurons using TriPure reagent (Roche) and purified using RNeasy MinElute Cleanup Kit (Promega) following standard protocols. .. The RNA concentration was measured by Nanodrop 2000 UV spectrophotometer and 1µg total RNA was converted to cDNA using the iScript Reverse Transcription Supermix (BioRad) system.

    Sequencing:

    Article Title: Upregulation of erythropoietin and erythropoietin receptor in castration-resistant progression of prostate cancer
    Article Snippet: .. For qRT-PCR analysis, cDNA was amplified with a SYBR Green PCR Kit (Roche) on an ABI PRISM 7900 sequence detection system, with the following EPO primers (sense 5'-ACC AAC ATT GCT TGT GCC AC-3' and antisense 5'-TCT GAA TGC TTC CTG CTC TGG-3') and EPOR primers (sense 5'-ACC GTG TCA TCC ACA TCA AT-3' and antisense 5'-GCC TTC AAA CTC GCT CTC TG-3'). .. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was amplified as an internal control.

    Staining:

    Article Title: Pea p68, a DEAD-box helicase, enhances salt tolerance in marker-free transgenic plants of soybean [Glycine max (L.) Merrill]
    Article Snippet: .. The amplified products were determined on a 1.0% agarose gel (Sigma, St. Louis, USA) stained with 0.1% ethidium bromide (Sigma, St. Louis, USA) and photographed using the Gel Documentation system (UVITEC, Cambridge, UK). qRT-PCR analysis was performed in LightCycler® 480 Real-time PCR system (Roche, Germany). .. The expression analysis of the RT-PCR positive (T1 ) and NT plants were conducted using Prime Script™ RT Reagent Kit (Takara Bio Inc, Japan) and p68 gene-specific primer set (FP: 5′-CCTCGCATTCTCTTCCTCGTA-3′ and RP: 5′-CGACGAGAACCATTGGCTAGA-3′) (Banu et al. ).

    Software:

    Article Title: Antimicrobial peptides and nitric oxide production by neutrophils from periodontitis subjects
    Article Snippet: .. The sequences of the primers for qRT-PCR analysis were obtained using the Light-Cycler software (Roche Diagnostics GmbH, Germany). .. The qRT-PCR assays were performed using the Light Cycler system (Roche Diagnostics GmbH).

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    Roche qrt pcr analysis total rna
    Quercetin increases uncoupling protein-1 ( Ucp1) gene expression specifically in subcutaneous white adipose tissue. Gene expression in sWAT ( A ) and BAT ( B ) was determined by <t>qRT-PCR.</t> Hematoxylin and eosin (H E) staining was performed on paraffin-embedded sWAT sections, and representative pictures are shown ( C ). Pictures were analyzed in ImageJ to determine the relative cell size ( D ). sWAT sections were stained for UCP-1 (arrows indicate UCP-1 positive cells, E ) as well. BAT sections were stained for H E ( F ) and used to quantify lipid droplet content in ImageJ ( G ). BAT sections were also stained for UCP-1 ( H ). Data are represented as mean ± SEM ( n = 8–10); the expression of genes was corrected for the reference gene β 2-microglobulin (sWAT), Gapdh , and Hprt (BAT), * p
    Qrt Pcr Analysis Total Rna, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche qrt pcr analysis
    Generating proliferative induced hepatocytes using defined transcription factors and oncogenic drivers ( A ) Schematic outline of the cell transformation assay for making lineage-specific cancer by lentiviral expression of three lineage-specific TFs to convert HFs to induced hepatocytes (iHep) and defined oncogenic drivers to transform iHeps to proliferating and tumorigenic cells. ( B ) Comparison of TF combinations ( Du et al , 2014 , Huang et al , 2014 , Morris et al , 2014 ) for converting human fibroblasts to iHeps by detecting transcript levels for liver marker genes ( ALBUMIN , TRANSFERRIN and SERPINA1/α-1-antitrypsin ) by <t>qRT-PCR</t> at different time points after iHep conversion, normalized to GAPDH levels (mean ± standard error).
    Qrt Pcr Analysis, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 296 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Roche quantitative real time pcr qrt pcr analysis qrt pcr analyses
    Features of miR-17-5p and ETV1 expression in TNBC cells and clinical samples. a , b , <t>QRT-PCR</t> analyses of miR-17-5p and ETV1 expression levels in TNBC cells. c , d , The expression levels of miR-17-5p and ETV1 in clinical TNBC samples evaluated by qRT-PCR. Data are expressed as the mean ± SEM of three independent experiments. ** P
    Quantitative Real Time Pcr Qrt Pcr Analysis Qrt Pcr Analyses, supplied by Roche, 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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    Image Search Results


    Quercetin increases uncoupling protein-1 ( Ucp1) gene expression specifically in subcutaneous white adipose tissue. Gene expression in sWAT ( A ) and BAT ( B ) was determined by qRT-PCR. Hematoxylin and eosin (H E) staining was performed on paraffin-embedded sWAT sections, and representative pictures are shown ( C ). Pictures were analyzed in ImageJ to determine the relative cell size ( D ). sWAT sections were stained for UCP-1 (arrows indicate UCP-1 positive cells, E ) as well. BAT sections were stained for H E ( F ) and used to quantify lipid droplet content in ImageJ ( G ). BAT sections were also stained for UCP-1 ( H ). Data are represented as mean ± SEM ( n = 8–10); the expression of genes was corrected for the reference gene β 2-microglobulin (sWAT), Gapdh , and Hprt (BAT), * p

    Journal: International Journal of Molecular Sciences

    Article Title: Quercetin Lowers Plasma Triglycerides Accompanied by White Adipose Tissue Browning in Diet-Induced Obese Mice

    doi: 10.3390/ijms19061786

    Figure Lengend Snippet: Quercetin increases uncoupling protein-1 ( Ucp1) gene expression specifically in subcutaneous white adipose tissue. Gene expression in sWAT ( A ) and BAT ( B ) was determined by qRT-PCR. Hematoxylin and eosin (H E) staining was performed on paraffin-embedded sWAT sections, and representative pictures are shown ( C ). Pictures were analyzed in ImageJ to determine the relative cell size ( D ). sWAT sections were stained for UCP-1 (arrows indicate UCP-1 positive cells, E ) as well. BAT sections were stained for H E ( F ) and used to quantify lipid droplet content in ImageJ ( G ). BAT sections were also stained for UCP-1 ( H ). Data are represented as mean ± SEM ( n = 8–10); the expression of genes was corrected for the reference gene β 2-microglobulin (sWAT), Gapdh , and Hprt (BAT), * p

    Article Snippet: RNA Isolation and qRT-PCR Analysis Total RNA was isolated using TriPure Isolation reagent (Roche obtained via Sigma, St. Louis, MO, USA ) following the manufacturer’s protocol. cDNA was made using Moloney Murine Leukemia Virus Reverse Transcriptase (Promega, Leiden, The Netherlands).

    Techniques: Expressing, Quantitative RT-PCR, Staining

    Quercetin reduces hepatic apolipoprotein B ( Apob) expression and increases the uptake of triglycerides (TG)-derived fatty acid (FA) by subcutaneous white adipose tissue. In week 2 and week 10 of the intervention, 24 h feces was collected ( A ) and used to determine fecal free fatty acid (FFA) concentration ( B ). Gene expression in the liver was determined by qRT-PCR for acyl-CoA synthetase long-chain family member 1 ( Acsl1) , acetyl-CoA carboxylase 2 ( Acc2 ), microsomal triglyceride transfer protein ( Mttp ), and Apob ( C ). After 12 weeks, mice were injected with glycerol tri[ 3 H]oleate-labeled lipoprotein-like particles, and clearance from plasma ( D ) and uptake per gram organ ( E ) were determined by 3 H-activity analysis. Data are represented as mean ± SEM ( n = 8–10); the expression of genes was corrected for the reference gene β2-microglobulin , * p

    Journal: International Journal of Molecular Sciences

    Article Title: Quercetin Lowers Plasma Triglycerides Accompanied by White Adipose Tissue Browning in Diet-Induced Obese Mice

    doi: 10.3390/ijms19061786

    Figure Lengend Snippet: Quercetin reduces hepatic apolipoprotein B ( Apob) expression and increases the uptake of triglycerides (TG)-derived fatty acid (FA) by subcutaneous white adipose tissue. In week 2 and week 10 of the intervention, 24 h feces was collected ( A ) and used to determine fecal free fatty acid (FFA) concentration ( B ). Gene expression in the liver was determined by qRT-PCR for acyl-CoA synthetase long-chain family member 1 ( Acsl1) , acetyl-CoA carboxylase 2 ( Acc2 ), microsomal triglyceride transfer protein ( Mttp ), and Apob ( C ). After 12 weeks, mice were injected with glycerol tri[ 3 H]oleate-labeled lipoprotein-like particles, and clearance from plasma ( D ) and uptake per gram organ ( E ) were determined by 3 H-activity analysis. Data are represented as mean ± SEM ( n = 8–10); the expression of genes was corrected for the reference gene β2-microglobulin , * p

    Article Snippet: RNA Isolation and qRT-PCR Analysis Total RNA was isolated using TriPure Isolation reagent (Roche obtained via Sigma, St. Louis, MO, USA ) following the manufacturer’s protocol. cDNA was made using Moloney Murine Leukemia Virus Reverse Transcriptase (Promega, Leiden, The Netherlands).

    Techniques: Expressing, Derivative Assay, Concentration Assay, Quantitative RT-PCR, Mouse Assay, Injection, Labeling, Activity Assay

    Generating proliferative induced hepatocytes using defined transcription factors and oncogenic drivers ( A ) Schematic outline of the cell transformation assay for making lineage-specific cancer by lentiviral expression of three lineage-specific TFs to convert HFs to induced hepatocytes (iHep) and defined oncogenic drivers to transform iHeps to proliferating and tumorigenic cells. ( B ) Comparison of TF combinations ( Du et al , 2014 , Huang et al , 2014 , Morris et al , 2014 ) for converting human fibroblasts to iHeps by detecting transcript levels for liver marker genes ( ALBUMIN , TRANSFERRIN and SERPINA1/α-1-antitrypsin ) by qRT-PCR at different time points after iHep conversion, normalized to GAPDH levels (mean ± standard error).

    Journal: bioRxiv

    Article Title: Cellular transformation by combined lineage conversion and oncogene expression

    doi: 10.1101/525600

    Figure Lengend Snippet: Generating proliferative induced hepatocytes using defined transcription factors and oncogenic drivers ( A ) Schematic outline of the cell transformation assay for making lineage-specific cancer by lentiviral expression of three lineage-specific TFs to convert HFs to induced hepatocytes (iHep) and defined oncogenic drivers to transform iHeps to proliferating and tumorigenic cells. ( B ) Comparison of TF combinations ( Du et al , 2014 , Huang et al , 2014 , Morris et al , 2014 ) for converting human fibroblasts to iHeps by detecting transcript levels for liver marker genes ( ALBUMIN , TRANSFERRIN and SERPINA1/α-1-antitrypsin ) by qRT-PCR at different time points after iHep conversion, normalized to GAPDH levels (mean ± standard error).

    Article Snippet: For qRT-PCR analysis, cDNA synthesis from two biological replicates was performed using the Transcriptor High-fidelity cDNA synthesis kit (Roche) and real-time PCR using SYBR green (Roche) with primers specific for each transcript (Table S2).

    Techniques: Cell Transformation Assay, Expressing, Marker, Quantitative RT-PCR

    SLC35B2 is required for endocytosis of α-Syn PFF (A) The CRISPR screen strategy. (B-D) SLC35B2 -KO cells are defective in endocytosis of α-Syn PFF but not monomeric α-Syn. (B) Validation of SLC35B2 (35B2)-KO cells by qRT-PCR. mRNA levels were normalized to that in control cells. Error bar, SEM, n=3. (C) Control, SLC35B2 -KO, or SLC35B2 -KO cells stably expressing FLAG-SLC35B2 were incubated with pHrodo-labeled α-Syn PFF (200nM for 4 h) and analyzed by FACS. FL, fluorescence. (D) Control and SLC35B2 -KO cells were treated with labeled α-Syn monomer at 800nM overnight prior to FACS analysis. (E-G) Knockdown (KD) of SLC35B2 attenuates α-Syn PFF uptake in primary neurons. (E) Primary neurons were infected with lentivirus expressing the indicated shRNAs together with EGFP (driven by the Synapsin promoter) at days-in- vitro 3 (DIV3). mRNA was purified from cells at DIV8 for qRT-PCR analysis. Error bar, SEM, n=2 biological repeats, each with triplicated PCR analyses. (F) Control or SLC35B2 -KD neurons expressing EGFP (green) at DIV8 were incubated with α-Syn PFF Alexa 594 (200nM) (red) for 4 h, stained with DAPI (blue), and analyzed by confocal microscopy. Scale bar, 5µm (G) Quantification of α-Syn PFF level in individual cells (indicated by dots) from two independent experiments. Error bar, SEM, p value from two-tailed t-test. A.U. arbitrary unit. (H, I) SLC35B2 is required for α-Syn PFF binding to the plasma membrane. ( H ) Control, SLC35B2 -KO, or SLC35B2 -KO cells re-expressing WT SLC35B2 were incubated with α-Syn PFF Alexa 594 (200nM) (magenta) on ice for 30min, stained with DAPI (blue), and imaged by confocal microscopy. Scale bar, 5µm. The graph in (I) shows the quantification of α-Syn PFF surface level in individual cells from two independent experiments. A.U. arbitrary unit.

    Journal: bioRxiv

    Article Title: A Myosin-7B dependent endocytosis pathway mediates cellular entry of α-Synuclein fibrils and polycation-bearing cargos

    doi: 10.1101/2020.02.12.946137

    Figure Lengend Snippet: SLC35B2 is required for endocytosis of α-Syn PFF (A) The CRISPR screen strategy. (B-D) SLC35B2 -KO cells are defective in endocytosis of α-Syn PFF but not monomeric α-Syn. (B) Validation of SLC35B2 (35B2)-KO cells by qRT-PCR. mRNA levels were normalized to that in control cells. Error bar, SEM, n=3. (C) Control, SLC35B2 -KO, or SLC35B2 -KO cells stably expressing FLAG-SLC35B2 were incubated with pHrodo-labeled α-Syn PFF (200nM for 4 h) and analyzed by FACS. FL, fluorescence. (D) Control and SLC35B2 -KO cells were treated with labeled α-Syn monomer at 800nM overnight prior to FACS analysis. (E-G) Knockdown (KD) of SLC35B2 attenuates α-Syn PFF uptake in primary neurons. (E) Primary neurons were infected with lentivirus expressing the indicated shRNAs together with EGFP (driven by the Synapsin promoter) at days-in- vitro 3 (DIV3). mRNA was purified from cells at DIV8 for qRT-PCR analysis. Error bar, SEM, n=2 biological repeats, each with triplicated PCR analyses. (F) Control or SLC35B2 -KD neurons expressing EGFP (green) at DIV8 were incubated with α-Syn PFF Alexa 594 (200nM) (red) for 4 h, stained with DAPI (blue), and analyzed by confocal microscopy. Scale bar, 5µm (G) Quantification of α-Syn PFF level in individual cells (indicated by dots) from two independent experiments. Error bar, SEM, p value from two-tailed t-test. A.U. arbitrary unit. (H, I) SLC35B2 is required for α-Syn PFF binding to the plasma membrane. ( H ) Control, SLC35B2 -KO, or SLC35B2 -KO cells re-expressing WT SLC35B2 were incubated with α-Syn PFF Alexa 594 (200nM) (magenta) on ice for 30min, stained with DAPI (blue), and imaged by confocal microscopy. Scale bar, 5µm. The graph in (I) shows the quantification of α-Syn PFF surface level in individual cells from two independent experiments. A.U. arbitrary unit.

    Article Snippet: qRT-PCR analysis of knockout or knockdown cells Total RNA was extracted from 3 million HEK293T cells or 0.75 million primary neurons using TriPure reagent (Roche) and purified using RNeasy MinElute Cleanup Kit (Promega) following standard protocols.

    Techniques: CRISPR, Quantitative RT-PCR, Stable Transfection, Expressing, Incubation, Labeling, FACS, Fluorescence, Infection, In Vitro, Purification, Polymerase Chain Reaction, Staining, Confocal Microscopy, Two Tailed Test, Binding Assay

    HSPG-mediated endocytosis requires MYO7B and actin filaments (A) Mapping the identified MYO7B sgRNA. (B) Validation of MYO7B -KO cells by qRT-PCR. mRNAs extracted from two clones of MYO7B KO cells and a control (Ctrl.) clone were analyzed by qRT-PCR. Error bars, SEM, n=3. (C) MYO7B KO cells are defective in α-Syn PFF uptake. Control (Ctrl.) and MYO7B -KO clones were incubated with pHrodo-labeled α-Syn PFF for 4 h before FACS analyses. (D) MYO7B is not required for endocytosis of α-Syn monomer. Control or MYO7B -KO cells were incubated with α-Syn monomer (800nM) overnight prior to FACS analysis. (E) MYO7B is not required for transferrin uptake. Control or MYO7B -KO cells were incubated with fluorescein-labeled transferrin (50µg/ml, 3h), washed, and analyzed with a fluorometer. Error bars, SEM, n=3. (F-H) KD of MYO7B in primary neurons reduces α-Syn PFF endocytosis without affecting its binding to the plasma membrane. (F) Primary neurons infected with the indicated shRNA-expressing lentivirus together with Synapsin_EGFP lentivirus at days-in- vitro 3 (DIV3) were incubated with α-Syn PFF Alexa 594 (200nM) for 4 h and imaged at DIV8. The insets show an enlarged view of the box. Arrows indicate peri-nuclear enrichment of α-Syn PFF positive vesicles in control cells. The graph in (G) shows the ratio of internalized α-Syn PFF relative to total α-Syn PFF in individual cells. (H) qRT-PCR evaluation of MYO7B mRNA levels using the same batch of cells. Error bars, SEM, n=two biological repeats with triplicated PCR analyses. (I-K) MYO7B dominant negative mutants inhibit α-Syn PFF uptake. (I) Representative confocal images of COS7 cells transfected with the indicated MYO7B plasmids and treated with α-Syn PFF Alexa 594 (400nM) for 3 h. Arrows indicate normal α-Syn PFF uptake in untransfected cells, which was used to normalize uptake. N, nuclei. (J) The graph shows the domain structure of the truncated MYO7B mutants. (K) Quantification of α-Syn PFF fluorescence (FL) in individual cells transfected (t) with the indicated MYO7B mutants normalized by the signal in untransfected cells (u).

    Journal: bioRxiv

    Article Title: A Myosin-7B dependent endocytosis pathway mediates cellular entry of α-Synuclein fibrils and polycation-bearing cargos

    doi: 10.1101/2020.02.12.946137

    Figure Lengend Snippet: HSPG-mediated endocytosis requires MYO7B and actin filaments (A) Mapping the identified MYO7B sgRNA. (B) Validation of MYO7B -KO cells by qRT-PCR. mRNAs extracted from two clones of MYO7B KO cells and a control (Ctrl.) clone were analyzed by qRT-PCR. Error bars, SEM, n=3. (C) MYO7B KO cells are defective in α-Syn PFF uptake. Control (Ctrl.) and MYO7B -KO clones were incubated with pHrodo-labeled α-Syn PFF for 4 h before FACS analyses. (D) MYO7B is not required for endocytosis of α-Syn monomer. Control or MYO7B -KO cells were incubated with α-Syn monomer (800nM) overnight prior to FACS analysis. (E) MYO7B is not required for transferrin uptake. Control or MYO7B -KO cells were incubated with fluorescein-labeled transferrin (50µg/ml, 3h), washed, and analyzed with a fluorometer. Error bars, SEM, n=3. (F-H) KD of MYO7B in primary neurons reduces α-Syn PFF endocytosis without affecting its binding to the plasma membrane. (F) Primary neurons infected with the indicated shRNA-expressing lentivirus together with Synapsin_EGFP lentivirus at days-in- vitro 3 (DIV3) were incubated with α-Syn PFF Alexa 594 (200nM) for 4 h and imaged at DIV8. The insets show an enlarged view of the box. Arrows indicate peri-nuclear enrichment of α-Syn PFF positive vesicles in control cells. The graph in (G) shows the ratio of internalized α-Syn PFF relative to total α-Syn PFF in individual cells. (H) qRT-PCR evaluation of MYO7B mRNA levels using the same batch of cells. Error bars, SEM, n=two biological repeats with triplicated PCR analyses. (I-K) MYO7B dominant negative mutants inhibit α-Syn PFF uptake. (I) Representative confocal images of COS7 cells transfected with the indicated MYO7B plasmids and treated with α-Syn PFF Alexa 594 (400nM) for 3 h. Arrows indicate normal α-Syn PFF uptake in untransfected cells, which was used to normalize uptake. N, nuclei. (J) The graph shows the domain structure of the truncated MYO7B mutants. (K) Quantification of α-Syn PFF fluorescence (FL) in individual cells transfected (t) with the indicated MYO7B mutants normalized by the signal in untransfected cells (u).

    Article Snippet: qRT-PCR analysis of knockout or knockdown cells Total RNA was extracted from 3 million HEK293T cells or 0.75 million primary neurons using TriPure reagent (Roche) and purified using RNeasy MinElute Cleanup Kit (Promega) following standard protocols.

    Techniques: Quantitative RT-PCR, Clone Assay, Incubation, Labeling, FACS, Binding Assay, Infection, shRNA, Expressing, In Vitro, Polymerase Chain Reaction, Dominant Negative Mutation, Transfection, Fluorescence

    Features of miR-17-5p and ETV1 expression in TNBC cells and clinical samples. a , b , QRT-PCR analyses of miR-17-5p and ETV1 expression levels in TNBC cells. c , d , The expression levels of miR-17-5p and ETV1 in clinical TNBC samples evaluated by qRT-PCR. Data are expressed as the mean ± SEM of three independent experiments. ** P

    Journal: BMC Cancer

    Article Title: miR-17-5p suppresses cell proliferation and invasion by targeting ETV1 in triple-negative breast cancer

    doi: 10.1186/s12885-017-3674-x

    Figure Lengend Snippet: Features of miR-17-5p and ETV1 expression in TNBC cells and clinical samples. a , b , QRT-PCR analyses of miR-17-5p and ETV1 expression levels in TNBC cells. c , d , The expression levels of miR-17-5p and ETV1 in clinical TNBC samples evaluated by qRT-PCR. Data are expressed as the mean ± SEM of three independent experiments. ** P

    Article Snippet: Quantitative real time PCR (qRT-PCR) analysis qRT-PCR analyses of miR-17-5p and ETV1 expression were performed on a LightCycler 480 (Roche Diagnostics, Germany) according to our previous report [ ].

    Techniques: Expressing, Quantitative RT-PCR

    ETV1 is a direct target of miR-17-5p in TNBC cells. a , Target sequences of miR-17-5p in ETV1 3′-UTR and mutant sites in 3′-UTR. b , Relative luciferase activity of ETV1 3′-UTR and mutant in the miR-17-5p mimic-transfected 293 T cells. c , d , The effect of miR-17-5p on ETV1 expression in MDA-MB-231 and BT549 cells was detected by qRT-PCR and western blotting after the cells were transfected with miR-17-5p mimic or inhibitor, respectively. e , The effect of miR-17-5p on ETV1 expression was also observed in MCF10A cells by co-transfecting with GV141-ETV1 and miR-17-5p inhibitor. Data are expressed as the mean ± SEM of three independent experiments. * P

    Journal: BMC Cancer

    Article Title: miR-17-5p suppresses cell proliferation and invasion by targeting ETV1 in triple-negative breast cancer

    doi: 10.1186/s12885-017-3674-x

    Figure Lengend Snippet: ETV1 is a direct target of miR-17-5p in TNBC cells. a , Target sequences of miR-17-5p in ETV1 3′-UTR and mutant sites in 3′-UTR. b , Relative luciferase activity of ETV1 3′-UTR and mutant in the miR-17-5p mimic-transfected 293 T cells. c , d , The effect of miR-17-5p on ETV1 expression in MDA-MB-231 and BT549 cells was detected by qRT-PCR and western blotting after the cells were transfected with miR-17-5p mimic or inhibitor, respectively. e , The effect of miR-17-5p on ETV1 expression was also observed in MCF10A cells by co-transfecting with GV141-ETV1 and miR-17-5p inhibitor. Data are expressed as the mean ± SEM of three independent experiments. * P

    Article Snippet: Quantitative real time PCR (qRT-PCR) analysis qRT-PCR analyses of miR-17-5p and ETV1 expression were performed on a LightCycler 480 (Roche Diagnostics, Germany) according to our previous report [ ].

    Techniques: Mutagenesis, Luciferase, Activity Assay, Transfection, Expressing, Multiple Displacement Amplification, Quantitative RT-PCR, Western Blot