quantitative real time polymerase chain reaction qrt pcr total rna  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99

    Structured Review

    Thermo Fisher quantitative real time polymerase chain reaction qrt pcr total rna
    MiR-21-5p directly targets SMAD7-3′-UTR in HMrSV5 cells. a Luciferase reporter assay results indicated that miR-21-5p directly bind to SMAD7 3’UTR. b Western blot was performed to measure the protein levels of SMAD7 in different groups. c <t>qRT-PCR</t> detection of SMAD7 mRNA in HMrSV5 cells transfected with miR-21-5p-mimics, mimics-NC, miR-21-5p-inhibitor and inhibitor-NC. d Western blot analysis for the protein levels of SMAD2/3, p-SMAD2/3, and TβRI in different groups. GAPDH was used as an internal reference. Each experiment was repeated at least three times. All the data were expressed as mean ± SEM (Student’s t -test * P
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr Total Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 364 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time polymerase chain reaction qrt pcr total rna/product/Thermo Fisher
    Average 99 stars, based on 364 article reviews
    Price from $9.99 to $1999.99
    quantitative real time polymerase chain reaction qrt pcr total rna - by Bioz Stars, 2020-04
    99/100 stars

    Images

    1) Product Images from "Exosomal miR-21-5p derived from gastric cancer promotes peritoneal metastasis via mesothelial-to-mesenchymal transition"

    Article Title: Exosomal miR-21-5p derived from gastric cancer promotes peritoneal metastasis via mesothelial-to-mesenchymal transition

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-018-0928-8

    MiR-21-5p directly targets SMAD7-3′-UTR in HMrSV5 cells. a Luciferase reporter assay results indicated that miR-21-5p directly bind to SMAD7 3’UTR. b Western blot was performed to measure the protein levels of SMAD7 in different groups. c qRT-PCR detection of SMAD7 mRNA in HMrSV5 cells transfected with miR-21-5p-mimics, mimics-NC, miR-21-5p-inhibitor and inhibitor-NC. d Western blot analysis for the protein levels of SMAD2/3, p-SMAD2/3, and TβRI in different groups. GAPDH was used as an internal reference. Each experiment was repeated at least three times. All the data were expressed as mean ± SEM (Student’s t -test * P
    Figure Legend Snippet: MiR-21-5p directly targets SMAD7-3′-UTR in HMrSV5 cells. a Luciferase reporter assay results indicated that miR-21-5p directly bind to SMAD7 3’UTR. b Western blot was performed to measure the protein levels of SMAD7 in different groups. c qRT-PCR detection of SMAD7 mRNA in HMrSV5 cells transfected with miR-21-5p-mimics, mimics-NC, miR-21-5p-inhibitor and inhibitor-NC. d Western blot analysis for the protein levels of SMAD2/3, p-SMAD2/3, and TβRI in different groups. GAPDH was used as an internal reference. Each experiment was repeated at least three times. All the data were expressed as mean ± SEM (Student’s t -test * P

    Techniques Used: Luciferase, Reporter Assay, Western Blot, Quantitative RT-PCR, Transfection

    MiR-21-5p induces MMT in HMrSV5 cells in vitro. a qRT-PCR detection of miR-21-5p expression in HMrSV5 transfected with miR-21-5p-mimics, mimics-NC, miR-21-5p-inhibitor and inhibitor-NC. Mimics-NC was the negative control for miR-21-5p-mimics and inhibitor-NC was the negative control for miR-21-5p-inhibitor. b, c Adhesion (Scale bar = 100 µm) and invasion (Scale bar = 100 µm) assays showed the effect of miR-21-5p on the HMrSV5 in vitro. d, e Western blot assays were performed to confirm the MMT of HMrSV5. GAPDH was used as an internal control. f Immunofluorescence assays were performed to confirm the MMT of HMrSV5. Scale bar = 50 µm. Each experiment was repeated at least three times. All the data were expressed as the mean ± SEM (Student’s t -test * P
    Figure Legend Snippet: MiR-21-5p induces MMT in HMrSV5 cells in vitro. a qRT-PCR detection of miR-21-5p expression in HMrSV5 transfected with miR-21-5p-mimics, mimics-NC, miR-21-5p-inhibitor and inhibitor-NC. Mimics-NC was the negative control for miR-21-5p-mimics and inhibitor-NC was the negative control for miR-21-5p-inhibitor. b, c Adhesion (Scale bar = 100 µm) and invasion (Scale bar = 100 µm) assays showed the effect of miR-21-5p on the HMrSV5 in vitro. d, e Western blot assays were performed to confirm the MMT of HMrSV5. GAPDH was used as an internal control. f Immunofluorescence assays were performed to confirm the MMT of HMrSV5. Scale bar = 50 µm. Each experiment was repeated at least three times. All the data were expressed as the mean ± SEM (Student’s t -test * P

    Techniques Used: In Vitro, Quantitative RT-PCR, Expressing, Transfection, Negative Control, Western Blot, Immunofluorescence

    GC-derived exosomes transfer miR-21-5p from donor cells to recipient cells. a The expression levels of miR-21-5p in TCGA dataset between GC tissues and normal tissues. All normalized expression values were expressed in log 2 ((RPM + 1)). b The expression levels of miR-21-5p in 49 paired GC tissues and adjacent normal tissues. All normalized expression values were expressed in log 2 2 −ΔΔCT . c qRT-PCR detection of miR-21-5p expression in exosomes derived from four GC cell lines and GES-1 cells. GES-1 cells-derived exosomes were used as the negative control. d qRT-PCR detection of miR-21-5p expression in HMrSV5 co-cultured with MGC803-derived exosomes and GES-1 cells-derived exosomes (negative control). e Exosomes with Cy3-labeled miR-21-5p or without Cy3-labeled miR-21-5p were added to HMrSV5 cells. MiR-21-5p without Cy3-label was used as the negative control. The fluorescence signals were detected under confocal microscope. Scale bar = 20 µm. f The levels of miR-21-5p in HMrSV5 cells transfected with miR-21-5p-NC or miR-21-inhibitor and in the related exosomes. g qRT-PCR detection of miR-21-5p expression in HMrSV5 cells co-cultured with exosomes derived from GES-1 cells or MGC803 cells transfected with miR-21-5p-NC or miR-21-5p-inhibitor. h Immunofluorescence assays were performed to measure the MMT protein markers (Vimentin and α-SMA). Scale bar = 50 µm. MiR-21-5p-NC group was used as the negative control. Each experiment was repeated at least three times. All the data were expressed as mean ± SEM, and the results of multiple comparisons were corrected with Bonferroni method (Student’s t -test * P
    Figure Legend Snippet: GC-derived exosomes transfer miR-21-5p from donor cells to recipient cells. a The expression levels of miR-21-5p in TCGA dataset between GC tissues and normal tissues. All normalized expression values were expressed in log 2 ((RPM + 1)). b The expression levels of miR-21-5p in 49 paired GC tissues and adjacent normal tissues. All normalized expression values were expressed in log 2 2 −ΔΔCT . c qRT-PCR detection of miR-21-5p expression in exosomes derived from four GC cell lines and GES-1 cells. GES-1 cells-derived exosomes were used as the negative control. d qRT-PCR detection of miR-21-5p expression in HMrSV5 co-cultured with MGC803-derived exosomes and GES-1 cells-derived exosomes (negative control). e Exosomes with Cy3-labeled miR-21-5p or without Cy3-labeled miR-21-5p were added to HMrSV5 cells. MiR-21-5p without Cy3-label was used as the negative control. The fluorescence signals were detected under confocal microscope. Scale bar = 20 µm. f The levels of miR-21-5p in HMrSV5 cells transfected with miR-21-5p-NC or miR-21-inhibitor and in the related exosomes. g qRT-PCR detection of miR-21-5p expression in HMrSV5 cells co-cultured with exosomes derived from GES-1 cells or MGC803 cells transfected with miR-21-5p-NC or miR-21-5p-inhibitor. h Immunofluorescence assays were performed to measure the MMT protein markers (Vimentin and α-SMA). Scale bar = 50 µm. MiR-21-5p-NC group was used as the negative control. Each experiment was repeated at least three times. All the data were expressed as mean ± SEM, and the results of multiple comparisons were corrected with Bonferroni method (Student’s t -test * P

    Techniques Used: Derivative Assay, Expressing, Quantitative RT-PCR, Negative Control, Cell Culture, Labeling, Fluorescence, Microscopy, Transfection, Immunofluorescence

    2) Product Images from "LINC00460 modulates KDM2A to promote cell proliferation and migration by targeting miR-342-3p in gastric cancer"

    Article Title: LINC00460 modulates KDM2A to promote cell proliferation and migration by targeting miR-342-3p in gastric cancer

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S169307

    LINC00460 up-regulates KDM2A expression by competitively binding to miR-342-3p. Notes: ( A ) MiR-342-3p expression was analyzed by qRT-PCR after transfection with LINC00460 mimics or si-LINC00460. ( B ) A putative binding site of miR-342-3p in the 3′UTR of KDM2A was predicted by miRanda online software. ( C ) Luciferase activity was analyzed after co-transfection of miR-342-3p and wild-type KDM2A 3′UTR or mutant KDM2A 3′UTR fragments. ( D ) KDM2A mRNA expression was analyzed by qRT-PCR after transfection with miR-NC or miR-342-3p mimics. ( E ) KDM2A protein expression was examined by Western blotting. ( F ) KDM2A mRNA expression was determined by qRT-PCR analysis after treatment with LINC00460 mimics or si-LINC00460. ( G ) Correlation between LINC00460 expression and miR-342-3p expression in GC tissues was determined by Pearson’s correlation analysis. ( H ) Correlation between LINC00460 expression and KDM2A mRNA expression in GC tissues was determined by Pearson’s correlation analysis. *** P
    Figure Legend Snippet: LINC00460 up-regulates KDM2A expression by competitively binding to miR-342-3p. Notes: ( A ) MiR-342-3p expression was analyzed by qRT-PCR after transfection with LINC00460 mimics or si-LINC00460. ( B ) A putative binding site of miR-342-3p in the 3′UTR of KDM2A was predicted by miRanda online software. ( C ) Luciferase activity was analyzed after co-transfection of miR-342-3p and wild-type KDM2A 3′UTR or mutant KDM2A 3′UTR fragments. ( D ) KDM2A mRNA expression was analyzed by qRT-PCR after transfection with miR-NC or miR-342-3p mimics. ( E ) KDM2A protein expression was examined by Western blotting. ( F ) KDM2A mRNA expression was determined by qRT-PCR analysis after treatment with LINC00460 mimics or si-LINC00460. ( G ) Correlation between LINC00460 expression and miR-342-3p expression in GC tissues was determined by Pearson’s correlation analysis. ( H ) Correlation between LINC00460 expression and KDM2A mRNA expression in GC tissues was determined by Pearson’s correlation analysis. *** P

    Techniques Used: Expressing, Binding Assay, Quantitative RT-PCR, Transfection, Software, Luciferase, Activity Assay, Cotransfection, Mutagenesis, Western Blot

    LINC00460 is highly expressed in GC tissues and cell lines. Notes: ( A ) LINC00460 expression data were downloaded from the TCGA database and analyzed. ( B ) LINC00460 expression in 60 pairs of GC tissues and adjacent normal tissues was analyzed by qRT-PCR. ( C ) LINC00460 expression in GC tissues and normal tissues were examined by ISH. ( D ) LINC00460 expression in one normal gastric epithelial cell line GSE1 and three GC cell lines was analyzed by qRT-PCR. ** P
    Figure Legend Snippet: LINC00460 is highly expressed in GC tissues and cell lines. Notes: ( A ) LINC00460 expression data were downloaded from the TCGA database and analyzed. ( B ) LINC00460 expression in 60 pairs of GC tissues and adjacent normal tissues was analyzed by qRT-PCR. ( C ) LINC00460 expression in GC tissues and normal tissues were examined by ISH. ( D ) LINC00460 expression in one normal gastric epithelial cell line GSE1 and three GC cell lines was analyzed by qRT-PCR. ** P

    Techniques Used: Expressing, Quantitative RT-PCR, In Situ Hybridization

    LINC00460 interacts with miR-342–3 p in GC cells. Notes: ( A ) A putative binding site of miR-342-3p in LINC00460 was predicted using the miRanda online software. ( B ) Luciferase activity was analyzed after co-transfection of miR-342-3p and wild-type LINC00460 and mutant LINC00460. ( C ) Anti-Ago2 RIP assays were performed to enrich the miRNAs interacting with LINC00460 in MGC803 cells after transfection with NC mimics or LINC00460 mimics, followed by qRT-PCR to examine the miR-342-3p levels in the immunoprecipitates. *** P
    Figure Legend Snippet: LINC00460 interacts with miR-342–3 p in GC cells. Notes: ( A ) A putative binding site of miR-342-3p in LINC00460 was predicted using the miRanda online software. ( B ) Luciferase activity was analyzed after co-transfection of miR-342-3p and wild-type LINC00460 and mutant LINC00460. ( C ) Anti-Ago2 RIP assays were performed to enrich the miRNAs interacting with LINC00460 in MGC803 cells after transfection with NC mimics or LINC00460 mimics, followed by qRT-PCR to examine the miR-342-3p levels in the immunoprecipitates. *** P

    Techniques Used: Binding Assay, Software, Luciferase, Activity Assay, Cotransfection, Mutagenesis, Transfection, Quantitative RT-PCR

    3) Product Images from "miR-96 exerts carcinogenic effect by activating AKT/GSK-3β/β-catenin signaling pathway through targeting inhibition of FOXO1 in hepatocellular carcinoma"

    Article Title: miR-96 exerts carcinogenic effect by activating AKT/GSK-3β/β-catenin signaling pathway through targeting inhibition of FOXO1 in hepatocellular carcinoma

    Journal: Cancer Cell International

    doi: 10.1186/s12935-019-0756-7

    Up-regulation of miR-96 and down-regulation of FOXO1 in tumor tissues and HepG2 cells. a miR-96 relative expression in tumor tissues and non-tumor tissues by qRT-PCR; b FOXO1 mRNA relative expression in tumor tissues and non-tumor tissues by qRT-PCR; c FOXO1 protein relative expression in tumor tissues and non-tumor tissues by Western blot; d miR-96 relative expression in L02 and HepG2 cells by qRT-PCR; e FOXO1 mRNA relative expression in L02 and HepG2 cells by qRT-PCR; f FOXO1 protein relative expression in L02 and HepG2 cells by Western blot. ** P
    Figure Legend Snippet: Up-regulation of miR-96 and down-regulation of FOXO1 in tumor tissues and HepG2 cells. a miR-96 relative expression in tumor tissues and non-tumor tissues by qRT-PCR; b FOXO1 mRNA relative expression in tumor tissues and non-tumor tissues by qRT-PCR; c FOXO1 protein relative expression in tumor tissues and non-tumor tissues by Western blot; d miR-96 relative expression in L02 and HepG2 cells by qRT-PCR; e FOXO1 mRNA relative expression in L02 and HepG2 cells by qRT-PCR; f FOXO1 protein relative expression in L02 and HepG2 cells by Western blot. ** P

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

    FOXO1 was directly suppressed by miR-96. a Prediction of binding sites for FOXO1 and miR-96 by Target Scan; b Luciferase reporter assay; c FOXO1 mRNA expression by qRT-PCR; d FOXO1 protein expression by Western blot. ## P
    Figure Legend Snippet: FOXO1 was directly suppressed by miR-96. a Prediction of binding sites for FOXO1 and miR-96 by Target Scan; b Luciferase reporter assay; c FOXO1 mRNA expression by qRT-PCR; d FOXO1 protein expression by Western blot. ## P

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

    4) Product Images from "Analysis of the miRNA Expression Profiles in the Zearalenone-Exposed TM3 Leydig Cell Line"

    Article Title: Analysis of the miRNA Expression Profiles in the Zearalenone-Exposed TM3 Leydig Cell Line

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20030635

    ( A ) Validation of the differentially expressed miRNAs. ( B ) Verification of the miRNA target genes. The qRT-PCR analyses of the mRNA expression levels ofmiR-146b-3p, miR-3098-5p, miR-185-3p, miR-467e-3p, miR-411-5p, miR-301a-5p, miR-210-5p, miR-326-3p, miR-615-5p, miR-410-3p, miR-96-3p, miR-19a-3p, miR-96-5p, miR-221-5p, miR-3057-5p, PTEN, AKT1, AKT2, AKT3, H-ras, N-ras, K-ras, Rap1a, Rap1b, Foxo1, Foxo3, Foxo4, and Foxo6 in the control and ZEA-exposed TM3 Leydig cells. The gene expression levels represent the mRNA expression levels relative to the control levels (the values represent mean ± SD). The asterisks are used to indicate a statistically significant difference: * p
    Figure Legend Snippet: ( A ) Validation of the differentially expressed miRNAs. ( B ) Verification of the miRNA target genes. The qRT-PCR analyses of the mRNA expression levels ofmiR-146b-3p, miR-3098-5p, miR-185-3p, miR-467e-3p, miR-411-5p, miR-301a-5p, miR-210-5p, miR-326-3p, miR-615-5p, miR-410-3p, miR-96-3p, miR-19a-3p, miR-96-5p, miR-221-5p, miR-3057-5p, PTEN, AKT1, AKT2, AKT3, H-ras, N-ras, K-ras, Rap1a, Rap1b, Foxo1, Foxo3, Foxo4, and Foxo6 in the control and ZEA-exposed TM3 Leydig cells. The gene expression levels represent the mRNA expression levels relative to the control levels (the values represent mean ± SD). The asterisks are used to indicate a statistically significant difference: * p

    Techniques Used: Quantitative RT-PCR, Expressing

    5) Product Images from "Targeting Gastrin-Releasing Peptide Suppresses Neuroblastoma Progression via Upregulation of PTEN Signaling"

    Article Title: Targeting Gastrin-Releasing Peptide Suppresses Neuroblastoma Progression via Upregulation of PTEN Signaling

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0072570

    GRP silencing on PTEN/AKT/mTOR signaling and MYCN, TWIST, FAK. (A) BE(2)-C/Tet/shGRP (+DOX) cells and SH-SY5Y/Tet/shGRP (+DOX) cells had an increase in PTEN expression along with correlative decreases in pAKT and pmTOR expression when compared to control cells (without doxycycline; -DOX). (B) Doxycycline-induced GRP silencing in BE (2)-C/Tet/shGRP cells decreased the mRNA levels of MYCN , TWIST and FAK by ∼50–60% as assessed by QRT-PCR (mean ± SEM; * = p
    Figure Legend Snippet: GRP silencing on PTEN/AKT/mTOR signaling and MYCN, TWIST, FAK. (A) BE(2)-C/Tet/shGRP (+DOX) cells and SH-SY5Y/Tet/shGRP (+DOX) cells had an increase in PTEN expression along with correlative decreases in pAKT and pmTOR expression when compared to control cells (without doxycycline; -DOX). (B) Doxycycline-induced GRP silencing in BE (2)-C/Tet/shGRP cells decreased the mRNA levels of MYCN , TWIST and FAK by ∼50–60% as assessed by QRT-PCR (mean ± SEM; * = p

    Techniques Used: Expressing, Quantitative RT-PCR

    6) Product Images from "Antinociceptive Effect of Tetrandrine on LPS-Induced Hyperalgesia via the Inhibition of IKK? Phosphorylation and the COX-2/PGE2 Pathway in Mice"

    Article Title: Antinociceptive Effect of Tetrandrine on LPS-Induced Hyperalgesia via the Inhibition of IKK? Phosphorylation and the COX-2/PGE2 Pathway in Mice

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0094586

    Repressive effects of TET on the expression of COX-1 and COX-2 in LPS-induced hyperalgesia. (A) Western blotting of COX-1 and COX-2 in brain tissues and the quantified comparison of relative densities are shown. (B) COX-1 and COX-2 mRNA expression were tested by qRT-PCR in brain tissues. (C) COX-1 and COX-2 mRNA expression were tested by qRT-PCR in cultured astroglia. Values are shown as M±SD, and normalized to the NS groups. *, P
    Figure Legend Snippet: Repressive effects of TET on the expression of COX-1 and COX-2 in LPS-induced hyperalgesia. (A) Western blotting of COX-1 and COX-2 in brain tissues and the quantified comparison of relative densities are shown. (B) COX-1 and COX-2 mRNA expression were tested by qRT-PCR in brain tissues. (C) COX-1 and COX-2 mRNA expression were tested by qRT-PCR in cultured astroglia. Values are shown as M±SD, and normalized to the NS groups. *, P

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

    7) Product Images from "Strigolactone analogues induce apoptosis through activation of p38 and the stress response pathway in cancer cell lines and in conditionally reprogramed primary prostate cancer cells."

    Article Title: Strigolactone analogues induce apoptosis through activation of p38 and the stress response pathway in cancer cell lines and in conditionally reprogramed primary prostate cancer cells.

    Journal: Oncotarget

    doi:

    Global changes in gene expression following treatment with ST362 or MEB55. A. U2OS cells were treated with vehicle or 5 ppm of ST362 or 5 ppm MEB55 for 6 hrs. Total RNA was extracted and analyzed by expression array. Results of two independent experiments are displayed as a Heatmap of genes differentially expressed in U2OS cells treated with MEB55 or with ST362 for 6 hrs. Genes that were ±2 fold p≤0.05 upregulated are shown in red and down regulated transcripts are shown in green. A representative list of individual genes at 6 hrs (±2 fold p≤0.05) is presented in Table 2 and at 24 hrs in Table S1 . B-H qRT-PCR analysis of representative genes in A. I. A protein network of top upregulated genes in response to MEB55 and ST362 was constructed using the Ingenuity software.
    Figure Legend Snippet: Global changes in gene expression following treatment with ST362 or MEB55. A. U2OS cells were treated with vehicle or 5 ppm of ST362 or 5 ppm MEB55 for 6 hrs. Total RNA was extracted and analyzed by expression array. Results of two independent experiments are displayed as a Heatmap of genes differentially expressed in U2OS cells treated with MEB55 or with ST362 for 6 hrs. Genes that were ±2 fold p≤0.05 upregulated are shown in red and down regulated transcripts are shown in green. A representative list of individual genes at 6 hrs (±2 fold p≤0.05) is presented in Table 2 and at 24 hrs in Table S1 . B-H qRT-PCR analysis of representative genes in A. I. A protein network of top upregulated genes in response to MEB55 and ST362 was constructed using the Ingenuity software.

    Techniques Used: Expressing, Quantitative RT-PCR, Construct, Software

    8) Product Images from "HSP90 inhibition downregulates thymidylate synthase and sensitizes colorectal cancer cell lines to the effect of 5FU-based chemotherapy"

    Article Title: HSP90 inhibition downregulates thymidylate synthase and sensitizes colorectal cancer cell lines to the effect of 5FU-based chemotherapy

    Journal: Oncotarget

    doi:

    Cell cycle effects of ganetespib in colorectal cancer cell lines. Inhibition of HSP90 decreases levels of Cdk4, cyclin D1, pRb, E2F1, and TS and increases p21 (A B) Cells were treated with vehicle (DMSO) or ganetespib (50nM) for 24 hours. Cell cycle arrest and the DNA content of CRC cells were measured by FACS analysis. Representative images from (a) HCT-116 and (B) HT-29 untreated and treated cells. (C) Cells were treated with vehicle (DMSO) or ganetespib (50nM) for 24 hours. Protein was extracted as described in the Methods section. Western blot analysis revealed increased expression of p21 and decreased expression of Cdk4, cyclin D1, pRb, E2F1, and TS in both cell lines treated with ganetespib. (D) Cells were treated with vehicle (DMSO) or ganetespib (50nM) for 24 hours. mRNA was extracted as described in the Methods section. mRNA was analyzed by qRT-PCR using primers for the indicated genes. Comparable qRT-PCR results were normalized with actin. Each value represents the mean ± standard deviation, obtained from determinations made on five cultures per experimental condition. qRT-PCR, quantitative real time-polymerase chain reaction. Ganetespib-treated HCT-116 and HT-29 cells showed significantly (*** p
    Figure Legend Snippet: Cell cycle effects of ganetespib in colorectal cancer cell lines. Inhibition of HSP90 decreases levels of Cdk4, cyclin D1, pRb, E2F1, and TS and increases p21 (A B) Cells were treated with vehicle (DMSO) or ganetespib (50nM) for 24 hours. Cell cycle arrest and the DNA content of CRC cells were measured by FACS analysis. Representative images from (a) HCT-116 and (B) HT-29 untreated and treated cells. (C) Cells were treated with vehicle (DMSO) or ganetespib (50nM) for 24 hours. Protein was extracted as described in the Methods section. Western blot analysis revealed increased expression of p21 and decreased expression of Cdk4, cyclin D1, pRb, E2F1, and TS in both cell lines treated with ganetespib. (D) Cells were treated with vehicle (DMSO) or ganetespib (50nM) for 24 hours. mRNA was extracted as described in the Methods section. mRNA was analyzed by qRT-PCR using primers for the indicated genes. Comparable qRT-PCR results were normalized with actin. Each value represents the mean ± standard deviation, obtained from determinations made on five cultures per experimental condition. qRT-PCR, quantitative real time-polymerase chain reaction. Ganetespib-treated HCT-116 and HT-29 cells showed significantly (*** p

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

    9) Product Images from "miR-22 inhibits mouse ovarian granulosa cell apoptosis by targeting SIRT1"

    Article Title: miR-22 inhibits mouse ovarian granulosa cell apoptosis by targeting SIRT1

    Journal: Biology Open

    doi: 10.1242/bio.016907

    The expression of miR-22 in mouse ovarian granulosa cells. The relative expression levels of miR-22 were measured in healthy follicles (HF), early atretic follicles (EAF), and progressively atretic follicles (PAF) using qRT-PCR. U6 was used as a loading control to normalize expression levels. Data were expressed as the mean±s.d. ( n =3). * P
    Figure Legend Snippet: The expression of miR-22 in mouse ovarian granulosa cells. The relative expression levels of miR-22 were measured in healthy follicles (HF), early atretic follicles (EAF), and progressively atretic follicles (PAF) using qRT-PCR. U6 was used as a loading control to normalize expression levels. Data were expressed as the mean±s.d. ( n =3). * P

    Techniques Used: Expressing, Quantitative RT-PCR

    10) Product Images from "Annexin A2 Coordinates STAT3 to Regulate the Invasion and Migration of Colorectal Cancer Cells In Vitro"

    Article Title: Annexin A2 Coordinates STAT3 to Regulate the Invasion and Migration of Colorectal Cancer Cells In Vitro

    Journal: Gastroenterology Research and Practice

    doi: 10.1155/2016/3521453

    Comparisons of the expression and the phosphorylation of Annexin A2 and STAT3 between the colorectal cancer (CRC) tissue and the paratumor tissue. (a) and (b) The expression of Annexin A2 (a) and STAT3 (b) in mRNA level was assayed in CRC tissues ( n = 35) and in paratumor tissues ( n = 35) by relative qRT-PCR analysis; (c) representative western blot analysis of the expression in protein level and the phosphorylation of Annexin A2 (Tyrosine 23) and STAT3 (Tyrosine 705) in CRC tissues ( n = 15) and in paratumor tissues ( n = 15); (d) relative level of Annexin A2 and STAT3 to β -actin in protein level; (e) relative Annexin A2 and STAT3 with phosphorylation to nonphosphorylated Annexin A2 and STAT3. ∗ and ∗∗ represented p
    Figure Legend Snippet: Comparisons of the expression and the phosphorylation of Annexin A2 and STAT3 between the colorectal cancer (CRC) tissue and the paratumor tissue. (a) and (b) The expression of Annexin A2 (a) and STAT3 (b) in mRNA level was assayed in CRC tissues ( n = 35) and in paratumor tissues ( n = 35) by relative qRT-PCR analysis; (c) representative western blot analysis of the expression in protein level and the phosphorylation of Annexin A2 (Tyrosine 23) and STAT3 (Tyrosine 705) in CRC tissues ( n = 15) and in paratumor tissues ( n = 15); (d) relative level of Annexin A2 and STAT3 to β -actin in protein level; (e) relative Annexin A2 and STAT3 with phosphorylation to nonphosphorylated Annexin A2 and STAT3. ∗ and ∗∗ represented p

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

    11) Product Images from "Resveratrol Exerts Dosage-Dependent Effects on the Self-Renewal and Neural Differentiation of hUC-MSCs"

    Article Title: Resveratrol Exerts Dosage-Dependent Effects on the Self-Renewal and Neural Differentiation of hUC-MSCs

    Journal: Molecules and Cells

    doi: 10.14348/molcells.2016.2345

    RES regulates the gene and protein expression of SIRT1, PCNA, p53, p21, and p16. (A) qRT-PCR results of the mRNA level of SIRT1, PCNA, p53, p21 and p16 in RES treated cells was normalized by housekeeping gene GAPDH correspondingly and quantified relative to that of the control. Data shown are the mean values of three repeats. Error bars represent standard deviation. * p
    Figure Legend Snippet: RES regulates the gene and protein expression of SIRT1, PCNA, p53, p21, and p16. (A) qRT-PCR results of the mRNA level of SIRT1, PCNA, p53, p21 and p16 in RES treated cells was normalized by housekeeping gene GAPDH correspondingly and quantified relative to that of the control. Data shown are the mean values of three repeats. Error bars represent standard deviation. * p

    Techniques Used: Expressing, Quantitative RT-PCR, Standard Deviation

    12) Product Images from "Protein phosphatase 2A-B55δ enhances chemotherapy sensitivity of human hepatocellular carcinoma under the regulation of microRNA-133b"

    Article Title: Protein phosphatase 2A-B55δ enhances chemotherapy sensitivity of human hepatocellular carcinoma under the regulation of microRNA-133b

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    doi: 10.1186/s13046-016-0341-z

    miR-133b participates in cell cycle regulation by targeting PPP2R2D . a - c After transfection with miNC or 133mi, HepG2 cells were treated with or without 2.5 μg/ml cDDP for 12 h. a miR-133b and PPP2R2D mRNA levels. b Protein levels of B55δ, Cyclin B1, and Cyclin E1. c Cell cycle distribution was analyzed by FCM. d - f HepG2 cells transfected with inNC or 133in were treated with or without cDDP. d qRT-PCR, e WB, and ( f ) FCM were conducted as before. The data are expressed as mean ± SD of three independent experiments. * P
    Figure Legend Snippet: miR-133b participates in cell cycle regulation by targeting PPP2R2D . a - c After transfection with miNC or 133mi, HepG2 cells were treated with or without 2.5 μg/ml cDDP for 12 h. a miR-133b and PPP2R2D mRNA levels. b Protein levels of B55δ, Cyclin B1, and Cyclin E1. c Cell cycle distribution was analyzed by FCM. d - f HepG2 cells transfected with inNC or 133in were treated with or without cDDP. d qRT-PCR, e WB, and ( f ) FCM were conducted as before. The data are expressed as mean ± SD of three independent experiments. * P

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

    13) Product Images from "Overexpression of Cullin7 is associated with hepatocellular carcinoma progression and pathogenesis"

    Article Title: Overexpression of Cullin7 is associated with hepatocellular carcinoma progression and pathogenesis

    Journal: BMC Cancer

    doi: 10.1186/s12885-017-3839-7

    Knockdown of Cullin7 inhibited the proliferation, migration, and invasion capacities of HCC cells. a Knockdown of Cullin7 in specific shRNA-transduced stable HepG2 cells. β-actin was used as a loading control. b The transfection efficiency of shCullin7 was analyzed by measuring transcript levels using qRT-PCR analyses in HepG2 cells. c Silencing endogenous Cullin7 inhibited cell growth as determined using MTT assays. d Silencing endogenous Cullin7 inhibited cell growth as determined using colony formation assays. Summary graphs are presented for the colony formation assay that is outlined. e HepG2 shCullin7 and control vector cells were subjected to a wound healing assay (left panels). The uncovered areas in the wound healing assays were quantified as a percentage of the original wound area. f HepG2 shCullin7 and control vector cells were subjected to Transwell migration (upper panels) and Matrigel invasion assays (lower panels). Quantification of migrated cells through the membrane and invaded cells through Matrigel of each cell line are shown as a proportion of the vector controls. Error bars represent the mean ± SD of three independent experiments
    Figure Legend Snippet: Knockdown of Cullin7 inhibited the proliferation, migration, and invasion capacities of HCC cells. a Knockdown of Cullin7 in specific shRNA-transduced stable HepG2 cells. β-actin was used as a loading control. b The transfection efficiency of shCullin7 was analyzed by measuring transcript levels using qRT-PCR analyses in HepG2 cells. c Silencing endogenous Cullin7 inhibited cell growth as determined using MTT assays. d Silencing endogenous Cullin7 inhibited cell growth as determined using colony formation assays. Summary graphs are presented for the colony formation assay that is outlined. e HepG2 shCullin7 and control vector cells were subjected to a wound healing assay (left panels). The uncovered areas in the wound healing assays were quantified as a percentage of the original wound area. f HepG2 shCullin7 and control vector cells were subjected to Transwell migration (upper panels) and Matrigel invasion assays (lower panels). Quantification of migrated cells through the membrane and invaded cells through Matrigel of each cell line are shown as a proportion of the vector controls. Error bars represent the mean ± SD of three independent experiments

    Techniques Used: Migration, shRNA, Transfection, Quantitative RT-PCR, MTT Assay, Colony Assay, Plasmid Preparation, Wound Healing Assay

    Upregulation of Cullin7 enhances the proliferation, migration, and invasion capacities of HCC cells. a Ectopic expression of Cullin7 in HepG2 cells analyzed by western blotting. β-actin was used as a loading control. b The transfection efficiency of Cullin7 was analyzed by measuring transcript levels using qRT-PCR analyses in HepG2 cells. c Cell proliferation after Cullin7 overexpression in cells was measured using MTT assays. d Colony formation assays show that upregulation of Cullin7 promotes cell growth, and the summary graphs are presented for the colony formation assay that is outlined. e HepG2 Cullin7 and control vector cells were subjected to a wound healing assay. f Overexpression of Cullin7 promoted cell invasion and migration as determined by Transwell migration and Matrigel invasion assays. Quantification of migrated cells through the membrane and invaded cells through Matrigel of each cell line are shown as a proportion of the vector controls. Error bars represent the mean ± SD of three independent experiments
    Figure Legend Snippet: Upregulation of Cullin7 enhances the proliferation, migration, and invasion capacities of HCC cells. a Ectopic expression of Cullin7 in HepG2 cells analyzed by western blotting. β-actin was used as a loading control. b The transfection efficiency of Cullin7 was analyzed by measuring transcript levels using qRT-PCR analyses in HepG2 cells. c Cell proliferation after Cullin7 overexpression in cells was measured using MTT assays. d Colony formation assays show that upregulation of Cullin7 promotes cell growth, and the summary graphs are presented for the colony formation assay that is outlined. e HepG2 Cullin7 and control vector cells were subjected to a wound healing assay. f Overexpression of Cullin7 promoted cell invasion and migration as determined by Transwell migration and Matrigel invasion assays. Quantification of migrated cells through the membrane and invaded cells through Matrigel of each cell line are shown as a proportion of the vector controls. Error bars represent the mean ± SD of three independent experiments

    Techniques Used: Migration, Expressing, Western Blot, Transfection, Quantitative RT-PCR, Over Expression, MTT Assay, Colony Assay, Plasmid Preparation, Wound Healing Assay

    14) Product Images from "Doxycycline Attenuates Leptospira-Induced IL-1β by Suppressing NLRP3 Inflammasome Priming"

    Article Title: Doxycycline Attenuates Leptospira-Induced IL-1β by Suppressing NLRP3 Inflammasome Priming

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2017.00857

    Doxycycline suppressed NLRP3 inflammasome priming with upregulation of the expression of the Na/Kβ1 subunit during leptospira infection. The expression of NLRP3 mRNA (A) in sonicated leptospira-infected J774A.1 cells was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). The mRNA levels of NLRP3 in J774A.1 cells were normalized to the expression of the housekeeping gene GAPDH. The expression of NLRP3 protein (B) , the activation of caspase-1, and the release of mature IL-1β in cell extracts (Lysate) (C,D) were analyzed by western blot. The β-Actin protein was used as a control. The expression of Na/Kβ1 subunit mRNA was analyzed by qRT-PCR in sonicated (E) and live (F) leptospira-infected J774A.1 cells. The mRNA levels of Na/Kβ1 subunit in J774A.1 cells were normalized to the expression of the housekeeping gene GAPDH. All bars show the mean ± SD of three independent experiments and analyzed by the one-way ANOVA. * p
    Figure Legend Snippet: Doxycycline suppressed NLRP3 inflammasome priming with upregulation of the expression of the Na/Kβ1 subunit during leptospira infection. The expression of NLRP3 mRNA (A) in sonicated leptospira-infected J774A.1 cells was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). The mRNA levels of NLRP3 in J774A.1 cells were normalized to the expression of the housekeeping gene GAPDH. The expression of NLRP3 protein (B) , the activation of caspase-1, and the release of mature IL-1β in cell extracts (Lysate) (C,D) were analyzed by western blot. The β-Actin protein was used as a control. The expression of Na/Kβ1 subunit mRNA was analyzed by qRT-PCR in sonicated (E) and live (F) leptospira-infected J774A.1 cells. The mRNA levels of Na/Kβ1 subunit in J774A.1 cells were normalized to the expression of the housekeeping gene GAPDH. All bars show the mean ± SD of three independent experiments and analyzed by the one-way ANOVA. * p

    Techniques Used: Expressing, Infection, Sonication, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Activation Assay, Western Blot

    15) Product Images from "Leucine-rich alpha-2-glycoprotein-1 is up-regulated in colorectal cancer and is a tumor promoter"

    Article Title: Leucine-rich alpha-2-glycoprotein-1 is up-regulated in colorectal cancer and is a tumor promoter

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S153375

    ( A ) LRG1 mRNA expression was examined by qRT-PCR in 240 human colorectal cancer tissues and their adjacent normal mucosa tissues. Notes: LRG1 mRNA expression was normalized to that of actin in each sample. LRG1 mRNA expression is up-regulated in CRC tissue compared with normal tissue. ( B ) LRG1 mRNA expression was examined by qRT-PCR in 240 human colon cancer tissues and their adjacent normal mucosa tissues. LRG1 mRNA expression was normalized to that of actin in each sample. LRG1 mRNA expression is up-regulated in stage IV human colorectal cancer tissues compared with the other 3 stages. ** P
    Figure Legend Snippet: ( A ) LRG1 mRNA expression was examined by qRT-PCR in 240 human colorectal cancer tissues and their adjacent normal mucosa tissues. Notes: LRG1 mRNA expression was normalized to that of actin in each sample. LRG1 mRNA expression is up-regulated in CRC tissue compared with normal tissue. ( B ) LRG1 mRNA expression was examined by qRT-PCR in 240 human colon cancer tissues and their adjacent normal mucosa tissues. LRG1 mRNA expression was normalized to that of actin in each sample. LRG1 mRNA expression is up-regulated in stage IV human colorectal cancer tissues compared with the other 3 stages. ** P

    Techniques Used: Expressing, Quantitative RT-PCR

    16) Product Images from "Correlation between Trop2 and amphiregulin coexpression and overall survival in gastric cancer"

    Article Title: Correlation between Trop2 and amphiregulin coexpression and overall survival in gastric cancer

    Journal: Cancer Medicine

    doi: 10.1002/cam4.1018

    Trop2 and AREG mRNA expression in 26 pairs GC tissue pairs. Trop2 and AREG mRNA expression was examined by qRT ‐ PCR and normalized to β ‐actin. T=GC tissues; N=matched tumor neighbor tissues.
    Figure Legend Snippet: Trop2 and AREG mRNA expression in 26 pairs GC tissue pairs. Trop2 and AREG mRNA expression was examined by qRT ‐ PCR and normalized to β ‐actin. T=GC tissues; N=matched tumor neighbor tissues.

    Techniques Used: Expressing, Quantitative RT-PCR

    17) Product Images from "Long noncoding RNA HEGBC promotes tumorigenesis and metastasis of gallbladder cancer via forming a positive feedback loop with IL-11/STAT3 signaling pathway"

    Article Title: Long noncoding RNA HEGBC promotes tumorigenesis and metastasis of gallbladder cancer via forming a positive feedback loop with IL-11/STAT3 signaling pathway

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    doi: 10.1186/s13046-018-0847-7

    Depletion of IL-11 attenuated the oncogenic roles of HEGBC in GBC cells. a The expression of IL-11 in HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using qRT-PCR. b Cell proliferation of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using Glo cell viability assay. c Cell proliferation of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using EdU incorporation assay. The red color represents EdU-positive cells. Scale bars, 100 μm. d HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA were treated with 5 ng/ml doxorubicin for 24 h, and then cell apoptosis was detected using TUNEL assay. e Cell migration of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using transwell assay. Scale bars, 100 μm. Results are shown as mean ± s.d. of 3 independent experiments. ** P
    Figure Legend Snippet: Depletion of IL-11 attenuated the oncogenic roles of HEGBC in GBC cells. a The expression of IL-11 in HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using qRT-PCR. b Cell proliferation of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using Glo cell viability assay. c Cell proliferation of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using EdU incorporation assay. The red color represents EdU-positive cells. Scale bars, 100 μm. d HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA were treated with 5 ng/ml doxorubicin for 24 h, and then cell apoptosis was detected using TUNEL assay. e Cell migration of HEGBC stably overexpressed and control NOZ cells transfected with IL-11 specific shRNA was detected using transwell assay. Scale bars, 100 μm. Results are shown as mean ± s.d. of 3 independent experiments. ** P

    Techniques Used: Expressing, Stable Transfection, Transfection, shRNA, Quantitative RT-PCR, Viability Assay, TUNEL Assay, Migration, Transwell Assay

    HEGBC bound to the promoter of IL11 and activated IL-11/STAT3 signaling pathway. a The distribution of HEGBC in the cytoplasmic and nuclear fractions of NOZ cells was detected using cytoplasmic and nuclear RNA isolation followed by qRT-PCR. U6 and β-actin were used as nuclear and cytoplasmic controls, respectively. b Schematic outline of the predicted binding site for HEGBC on the promoter of IL11 . c ChIRP assay in NOZ cells was performed with antisense probe sets against HEGBC or LacZ (negative control). The bound DNA was detected using qRT-PCR with specific primers against the promoters of IL11 or ACTB . d The expression of IL-11 in HEGBC stably overexpressed and control NOZ cells was detected using qRT-PCR. e The expression of IL-11 in HEGBC stably depleted and control EH-GB2 cells was detected using qRT-PCR. f The concentration of IL11 in the culture medium from HEGBC stably overexpressed and control NOZ cells was detected using ELISA. g The concentration of IL11 in the culture medium from HEGBC stably depleted and control EH-GB2 cells was detected using ELISA. h STAT3 phosphorylation level in HEGBC stably overexpressed and control NOZ cells was detected using western blot. i STAT3 phosphorylation level in HEGBC stably depleted and control EH-GB2 cells was detected using western blot. j The expression of the target genes of STAT3 in HEGBC stably overexpressed and control NOZ cells was detected using qRT-PCR. k The expression of the target genes of STAT3 in HEGBC stably depleted and control EH-GB2 cells was detected using qRT-PCR. Results are shown as mean ± s.d. of 3 independent experiments. ** P
    Figure Legend Snippet: HEGBC bound to the promoter of IL11 and activated IL-11/STAT3 signaling pathway. a The distribution of HEGBC in the cytoplasmic and nuclear fractions of NOZ cells was detected using cytoplasmic and nuclear RNA isolation followed by qRT-PCR. U6 and β-actin were used as nuclear and cytoplasmic controls, respectively. b Schematic outline of the predicted binding site for HEGBC on the promoter of IL11 . c ChIRP assay in NOZ cells was performed with antisense probe sets against HEGBC or LacZ (negative control). The bound DNA was detected using qRT-PCR with specific primers against the promoters of IL11 or ACTB . d The expression of IL-11 in HEGBC stably overexpressed and control NOZ cells was detected using qRT-PCR. e The expression of IL-11 in HEGBC stably depleted and control EH-GB2 cells was detected using qRT-PCR. f The concentration of IL11 in the culture medium from HEGBC stably overexpressed and control NOZ cells was detected using ELISA. g The concentration of IL11 in the culture medium from HEGBC stably depleted and control EH-GB2 cells was detected using ELISA. h STAT3 phosphorylation level in HEGBC stably overexpressed and control NOZ cells was detected using western blot. i STAT3 phosphorylation level in HEGBC stably depleted and control EH-GB2 cells was detected using western blot. j The expression of the target genes of STAT3 in HEGBC stably overexpressed and control NOZ cells was detected using qRT-PCR. k The expression of the target genes of STAT3 in HEGBC stably depleted and control EH-GB2 cells was detected using qRT-PCR. Results are shown as mean ± s.d. of 3 independent experiments. ** P

    Techniques Used: Isolation, Quantitative RT-PCR, Binding Assay, Negative Control, Expressing, Stable Transfection, Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot

    p-STAT3 bound to the promoter of HEGBC and activated HEGBC expression. a Schematic outline of the predicted binding site for p-STAT3 on the promoter of HEGBC . b ChIP assay in NOZ cells was performed using p-STAT3 specific antibody or negative control IgG. The bound DNA was detected using qRT-PCR with specific primers against the predicted binding sites on the promoter of HEGBC or a distal non-binding site (negative control, NC). c RIP assay in NOZ cells was performed using p-STAT3 specific antibody, STAT3 specific antibody, or negative control IgG. The bound RNA was detected using qRT-PCR with specific primers against HEGBC or TSLNC8 (positive control). d After co-transfection of renilla luciferase reporter pRL-TK with firefly luciferase reporters containing HEGBC promoter, p-STAT3 binding sites mutated HEGBC promoter, or nothing into NOZ cells, the NOZ cells were treated with 5 μM p-STAT3 inhibitor SC144 for 72 h. Luciferase activities in these NOZ cells were detected. Data are presented as the relative ratio of firefly luciferase activity to renilla luciferase activity. e After co-transfection of renilla luciferase reporter pRL-TK with firefly luciferase reporters containing HEGBC promoter, p-STAT3 binding sites mutated HEGBC promoter, or nothing into NOZ cells, the NOZ cells were treated with 20 ng/mL IL-11 for 72 h. Luciferase activities in these NOZ cells were detected. Data are presented as the relative ratio of firefly luciferase activity to renilla luciferase activity. f The expression of IL-11 in NOZ cells treated with 5 μM SC144 for 72 h was detected using qRT-PCR. g The expression of IL-11 in NOZ cells treated with 20 ng/mL IL-11 for 72 h was detected using qRT-PCR. For b - g , results are shown as mean ± s.d. of 3 independent experiments. ns, not significant, ** P
    Figure Legend Snippet: p-STAT3 bound to the promoter of HEGBC and activated HEGBC expression. a Schematic outline of the predicted binding site for p-STAT3 on the promoter of HEGBC . b ChIP assay in NOZ cells was performed using p-STAT3 specific antibody or negative control IgG. The bound DNA was detected using qRT-PCR with specific primers against the predicted binding sites on the promoter of HEGBC or a distal non-binding site (negative control, NC). c RIP assay in NOZ cells was performed using p-STAT3 specific antibody, STAT3 specific antibody, or negative control IgG. The bound RNA was detected using qRT-PCR with specific primers against HEGBC or TSLNC8 (positive control). d After co-transfection of renilla luciferase reporter pRL-TK with firefly luciferase reporters containing HEGBC promoter, p-STAT3 binding sites mutated HEGBC promoter, or nothing into NOZ cells, the NOZ cells were treated with 5 μM p-STAT3 inhibitor SC144 for 72 h. Luciferase activities in these NOZ cells were detected. Data are presented as the relative ratio of firefly luciferase activity to renilla luciferase activity. e After co-transfection of renilla luciferase reporter pRL-TK with firefly luciferase reporters containing HEGBC promoter, p-STAT3 binding sites mutated HEGBC promoter, or nothing into NOZ cells, the NOZ cells were treated with 20 ng/mL IL-11 for 72 h. Luciferase activities in these NOZ cells were detected. Data are presented as the relative ratio of firefly luciferase activity to renilla luciferase activity. f The expression of IL-11 in NOZ cells treated with 5 μM SC144 for 72 h was detected using qRT-PCR. g The expression of IL-11 in NOZ cells treated with 20 ng/mL IL-11 for 72 h was detected using qRT-PCR. For b - g , results are shown as mean ± s.d. of 3 independent experiments. ns, not significant, ** P

    Techniques Used: Expressing, Binding Assay, Chromatin Immunoprecipitation, Negative Control, Quantitative RT-PCR, Positive Control, Cotransfection, Luciferase, Activity Assay

    HEGBC was up-regulated in GBC and associated with poor survival of GBC patients. a The expression of HEGBC in 102 pairs of GBC tissues and adjacent non-tumor tissues was detected using qRT-PCR. P
    Figure Legend Snippet: HEGBC was up-regulated in GBC and associated with poor survival of GBC patients. a The expression of HEGBC in 102 pairs of GBC tissues and adjacent non-tumor tissues was detected using qRT-PCR. P

    Techniques Used: Expressing, Quantitative RT-PCR

    Ectopic expression of HEGBC promoted GBC cell proliferation and migration in vitro. a The expression of HEGBC in HEGBC stably overexpressed and control SGC-996 cells was detected using qRT-PCR. b The expression of HEGBC in HEGBC stably overexpressed and control NOZ cells was detected using qRT-PCR. c Cell proliferation of HEGBC stably overexpressed and control SGC-996 cells was detected using Glo cell viability assay. d Cell proliferation of HEGBC stably overexpressed and control NOZ cells was detected using Glo cell viability assay. e Cell proliferation of HEGBC stably overexpressed and control SGC-996 and NOZ cells was detected using EdU incorporation assay. The red color represents EdU-positive cells. Scale bars, 100 μm. f HEGBC stably overexpressed and control SGC-996 and NOZ cells were treated with 5 ng/mL doxorubicin for 24 h, and then cell apoptosis was detected using TUNEL assay. g Cell migration of HEGBC stably overexpressed and control SGC-996 and NOZ cells was detected using transwell assay. Scale bars, 100 μm. Results are shown as mean ± s.d. of 3 independent experiments. * P
    Figure Legend Snippet: Ectopic expression of HEGBC promoted GBC cell proliferation and migration in vitro. a The expression of HEGBC in HEGBC stably overexpressed and control SGC-996 cells was detected using qRT-PCR. b The expression of HEGBC in HEGBC stably overexpressed and control NOZ cells was detected using qRT-PCR. c Cell proliferation of HEGBC stably overexpressed and control SGC-996 cells was detected using Glo cell viability assay. d Cell proliferation of HEGBC stably overexpressed and control NOZ cells was detected using Glo cell viability assay. e Cell proliferation of HEGBC stably overexpressed and control SGC-996 and NOZ cells was detected using EdU incorporation assay. The red color represents EdU-positive cells. Scale bars, 100 μm. f HEGBC stably overexpressed and control SGC-996 and NOZ cells were treated with 5 ng/mL doxorubicin for 24 h, and then cell apoptosis was detected using TUNEL assay. g Cell migration of HEGBC stably overexpressed and control SGC-996 and NOZ cells was detected using transwell assay. Scale bars, 100 μm. Results are shown as mean ± s.d. of 3 independent experiments. * P

    Techniques Used: Expressing, Migration, In Vitro, Stable Transfection, Quantitative RT-PCR, Viability Assay, TUNEL Assay, Transwell Assay

    Depletion of HEGBC inhibited GBC cell proliferation and migration in vitro. a The expression of HEGBC in HEGBC stably depleted and control GBC-SD cells was detected using qRT-PCR. b The expression of HEGBC in HEGBC stably depleted and control EH-GB2 cells was detected using qRT-PCR. c Cell proliferation of HEGBC stably depleted and control GBC-SD cells was detected using Glo cell viability assay. d Cell proliferation of HEGBC stably depleted and control EH-GB2 cells was detected using Glo cell viability assay. e Cell proliferation of HEGBC stably depleted and control GBC-SD and EH-GB2 cells was detected using EdU incorporation assay. The red color represents EdU-positive cells. Scale bars, 100 μm. f HEGBC stably depleted and control GBC-SD and EH-GB2 cells were treated with 5 ng/mL doxorubicin for 24 h, and then cell apoptosis was detected using TUNEL assay. g Cell migration of HEGBC stably depleted and control GBC-SD and EH-GB2 cells was detected using transwell assay. Scale bars, 100 μm. Results are shown as mean ± s.d. of 3 independent experiments. * P
    Figure Legend Snippet: Depletion of HEGBC inhibited GBC cell proliferation and migration in vitro. a The expression of HEGBC in HEGBC stably depleted and control GBC-SD cells was detected using qRT-PCR. b The expression of HEGBC in HEGBC stably depleted and control EH-GB2 cells was detected using qRT-PCR. c Cell proliferation of HEGBC stably depleted and control GBC-SD cells was detected using Glo cell viability assay. d Cell proliferation of HEGBC stably depleted and control EH-GB2 cells was detected using Glo cell viability assay. e Cell proliferation of HEGBC stably depleted and control GBC-SD and EH-GB2 cells was detected using EdU incorporation assay. The red color represents EdU-positive cells. Scale bars, 100 μm. f HEGBC stably depleted and control GBC-SD and EH-GB2 cells were treated with 5 ng/mL doxorubicin for 24 h, and then cell apoptosis was detected using TUNEL assay. g Cell migration of HEGBC stably depleted and control GBC-SD and EH-GB2 cells was detected using transwell assay. Scale bars, 100 μm. Results are shown as mean ± s.d. of 3 independent experiments. * P

    Techniques Used: Migration, In Vitro, Expressing, Stable Transfection, Quantitative RT-PCR, Viability Assay, TUNEL Assay, Transwell Assay

    18) Product Images from "G3BP1 inhibits RNA virus replication by positively regulating RIG-I-mediated cellular antiviral response"

    Article Title: G3BP1 inhibits RNA virus replication by positively regulating RIG-I-mediated cellular antiviral response

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-019-2178-9

    G3BP1 positively regulates the cellular antiviral response. a , b G3BP1-overexpressed HEK293T cell lines a or G3BP1-deficient HEK293T cells b were infected with SeV or VSV-GFP (MOI = 0.1) for the indicated time, and then the cell lysates were analyzed by immunoblotting with the antibodies against SeV, GFP, or β-actin. c Effects of G3BP1 on SeV and VSV infection. G3BP1-overexpressed or G3BP1-deficient and control HEK293T cells were infected with SeV for 12 h or with VSV-GFP (MOI = 0.1) for 4 h. The mRNA level of the SeV P and VSV P proteins in cells was determined by qRT-PCR. The experiment was repeated in triplicates. d Effects of G3BP1-overexpressed on VSV titer. G3BP1-overexpressed HEK293T cells were transfected with 1 μg/ml poly (I:C) for 16 h and infected with VSV-GFP (MOI = 0.1) for 18 h. Supernatants were then analyzed for VSV production by standard plaque assays. The experiment was repeated in triplicates. e G3BP1-overexpressed HEK293T cells were infected with VSV-GFP (MOI = 0.1) for 4 h. Images were captured by fluorescence microscopy. In addition, the GFP fluorescence levels in VSV-GFP-infected cells were analyzed by flow cytometry. The experiment was repeated in triplicates. f Effects of G3BP1-deficient on VSV titer. The experiments were similarly to those described in c . The experiment was repeated in triplicates. g G3BP1-deficient HEK293T cells were infected with VSV-GFP (MOI = 0.1) for 2 h. Images were then captured by fluorescence microscopy. In addition, the GFP fluorescence levels in VSV-GFP-infected cells were analyzed by flow cytometry. The experiment was repeated in triplicates. qRT-PCR, quantitative real-time polymerase chain reaction. The experiments were similarly described in b . Data are mean ± SD of three independent experiments. * P
    Figure Legend Snippet: G3BP1 positively regulates the cellular antiviral response. a , b G3BP1-overexpressed HEK293T cell lines a or G3BP1-deficient HEK293T cells b were infected with SeV or VSV-GFP (MOI = 0.1) for the indicated time, and then the cell lysates were analyzed by immunoblotting with the antibodies against SeV, GFP, or β-actin. c Effects of G3BP1 on SeV and VSV infection. G3BP1-overexpressed or G3BP1-deficient and control HEK293T cells were infected with SeV for 12 h or with VSV-GFP (MOI = 0.1) for 4 h. The mRNA level of the SeV P and VSV P proteins in cells was determined by qRT-PCR. The experiment was repeated in triplicates. d Effects of G3BP1-overexpressed on VSV titer. G3BP1-overexpressed HEK293T cells were transfected with 1 μg/ml poly (I:C) for 16 h and infected with VSV-GFP (MOI = 0.1) for 18 h. Supernatants were then analyzed for VSV production by standard plaque assays. The experiment was repeated in triplicates. e G3BP1-overexpressed HEK293T cells were infected with VSV-GFP (MOI = 0.1) for 4 h. Images were captured by fluorescence microscopy. In addition, the GFP fluorescence levels in VSV-GFP-infected cells were analyzed by flow cytometry. The experiment was repeated in triplicates. f Effects of G3BP1-deficient on VSV titer. The experiments were similarly to those described in c . The experiment was repeated in triplicates. g G3BP1-deficient HEK293T cells were infected with VSV-GFP (MOI = 0.1) for 2 h. Images were then captured by fluorescence microscopy. In addition, the GFP fluorescence levels in VSV-GFP-infected cells were analyzed by flow cytometry. The experiment was repeated in triplicates. qRT-PCR, quantitative real-time polymerase chain reaction. The experiments were similarly described in b . Data are mean ± SD of three independent experiments. * P

    Techniques Used: Infection, Quantitative RT-PCR, Transfection, Fluorescence, Microscopy, Flow Cytometry, Cytometry, Real-time Polymerase Chain Reaction

    19) Product Images from "Induced Pluripotency of Human Prostatic Epithelial Cells"

    Article Title: Induced Pluripotency of Human Prostatic Epithelial Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0064503

    Determination of expression levels of pluripotent genes and methylation levels of their promoters in E-PZ-1 and E-PZ-1-iPS-like cells. mRNA levels of Nanog (A), Rex1 (B), Oct4 (C), Klf4 (D), c-Myc (E), Sox2 (F), and CD133 (G) were measured by qRT-PCR and normalized against TBP. Methylation of Nanog (H) and Oct4 (I) promoters were determined by bisulfite pyrosequencing. The Y-axis is the fold-level of gene expression or promoter methylation in E-PZ-1-iPS-like cells compared to those in E-PZ-1 cells, which were set as 1. (J) and (K) were histograms of the number of chromosomes in 100 E-PZ-1-iPS-4 and -7 cells, respectively, determined by metaphase chromosome counting.
    Figure Legend Snippet: Determination of expression levels of pluripotent genes and methylation levels of their promoters in E-PZ-1 and E-PZ-1-iPS-like cells. mRNA levels of Nanog (A), Rex1 (B), Oct4 (C), Klf4 (D), c-Myc (E), Sox2 (F), and CD133 (G) were measured by qRT-PCR and normalized against TBP. Methylation of Nanog (H) and Oct4 (I) promoters were determined by bisulfite pyrosequencing. The Y-axis is the fold-level of gene expression or promoter methylation in E-PZ-1-iPS-like cells compared to those in E-PZ-1 cells, which were set as 1. (J) and (K) were histograms of the number of chromosomes in 100 E-PZ-1-iPS-4 and -7 cells, respectively, determined by metaphase chromosome counting.

    Techniques Used: Expressing, Methylation, Quantitative RT-PCR

    20) Product Images from "Hypoxia-Inducible MiR-210 Is an Independent Prognostic Factor and Contributes to Metastasis in Colorectal Cancer"

    Article Title: Hypoxia-Inducible MiR-210 Is an Independent Prognostic Factor and Contributes to Metastasis in Colorectal Cancer

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0090952

    MiR-210 suppresses VMP1 expression by directly binding to its 3′-UTR. (A) The putative miR-210 binding sequences in VMP1 3′-UTR. (B) Luciferase activity assay was performed for the HEK293T cells cotransfected with pmiR-REPORT™ vectors containing WT-VMP1 3′-UTR or MUT-VMP1 3′-UTR sequences and miR-210 mimics. Data are presented as normalized fold change in luciferase activity. (C, D) VMP1 mRNA and protein were determined in HT-29 cells and SW480 cells transfected with miR-210 mimics or miR-negative control by qRT-PCR and Western blot, respectively. (E) Inverse correlation between miR-210 expression and VMP1 mRNA levels in CRC tissues was analyzed using Pearson's correlation analysis.
    Figure Legend Snippet: MiR-210 suppresses VMP1 expression by directly binding to its 3′-UTR. (A) The putative miR-210 binding sequences in VMP1 3′-UTR. (B) Luciferase activity assay was performed for the HEK293T cells cotransfected with pmiR-REPORT™ vectors containing WT-VMP1 3′-UTR or MUT-VMP1 3′-UTR sequences and miR-210 mimics. Data are presented as normalized fold change in luciferase activity. (C, D) VMP1 mRNA and protein were determined in HT-29 cells and SW480 cells transfected with miR-210 mimics or miR-negative control by qRT-PCR and Western blot, respectively. (E) Inverse correlation between miR-210 expression and VMP1 mRNA levels in CRC tissues was analyzed using Pearson's correlation analysis.

    Techniques Used: Expressing, Binding Assay, Luciferase, Activity Assay, Transfection, Negative Control, Quantitative RT-PCR, Western Blot

    MiR-210 expression is increased in CRC tissues and associated with a poor prognosis. (A) MiR-210 expression was tested using qRT-PCR in 193 pairs of human CRC tissues (CRC) and adjacent non-tumorous tissues (NT), and its expression was normalized to the level of U6 small nuclear RNA (U6) expression in each sample. (B) Fold changes in miR-210 expression in each paired sample. The data are represented as a log 2 -fold change (cancer/normal), which was defined as > 1 (overexpression) and
    Figure Legend Snippet: MiR-210 expression is increased in CRC tissues and associated with a poor prognosis. (A) MiR-210 expression was tested using qRT-PCR in 193 pairs of human CRC tissues (CRC) and adjacent non-tumorous tissues (NT), and its expression was normalized to the level of U6 small nuclear RNA (U6) expression in each sample. (B) Fold changes in miR-210 expression in each paired sample. The data are represented as a log 2 -fold change (cancer/normal), which was defined as > 1 (overexpression) and

    Techniques Used: Expressing, Quantitative RT-PCR, Over Expression

    21) Product Images from "In Vitro Antileukemia Activity of ZSTK474 on K562 and Multidrug Resistant K562/A02 Cells"

    Article Title: In Vitro Antileukemia Activity of ZSTK474 on K562 and Multidrug Resistant K562/A02 Cells

    Journal: International Journal of Biological Sciences

    doi: 10.7150/ijbs.14878

    ZSTK474 affected the cell cycle-related proteins in K562 and K562/A02 cells. (A) Western blot analysis of the cell cycle-related proteins. K562 and K562/A02 cells were treated with indicated concentrations of ZSTK474 for 48 h. The levels of cyclin D1, p27, and p-pRb in nucleus were determined by western blot. (B) qRT-PCR analysis of p27 expression at mRNA level. Total RNA of the cells was extracted using TriZol reagent and quantified by a Nanodrop spectrophotometer. Five hundred ng of RNA was reverse-transcribed to cDNA. The PCR reaction was conducted in a 20 µl system containing 100 ng cDNA, 200 nM of former primer and reverse primer, using CFX96 TM Real-Time PCR Detection System. GAPDH was used as a housekeeping gene to normalize RNA expression. The relative gene expression levels were quantified by using the comparative Ct (ΔΔCt) method. Results represent mean ± SD of three independent experiments. *p
    Figure Legend Snippet: ZSTK474 affected the cell cycle-related proteins in K562 and K562/A02 cells. (A) Western blot analysis of the cell cycle-related proteins. K562 and K562/A02 cells were treated with indicated concentrations of ZSTK474 for 48 h. The levels of cyclin D1, p27, and p-pRb in nucleus were determined by western blot. (B) qRT-PCR analysis of p27 expression at mRNA level. Total RNA of the cells was extracted using TriZol reagent and quantified by a Nanodrop spectrophotometer. Five hundred ng of RNA was reverse-transcribed to cDNA. The PCR reaction was conducted in a 20 µl system containing 100 ng cDNA, 200 nM of former primer and reverse primer, using CFX96 TM Real-Time PCR Detection System. GAPDH was used as a housekeeping gene to normalize RNA expression. The relative gene expression levels were quantified by using the comparative Ct (ΔΔCt) method. Results represent mean ± SD of three independent experiments. *p

    Techniques Used: Western Blot, Quantitative RT-PCR, Expressing, Spectrophotometry, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, RNA Expression

    22) Product Images from "Expression and significance of histone H3K27 demethylases in renal cell carcinoma"

    Article Title: Expression and significance of histone H3K27 demethylases in renal cell carcinoma

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-12-470

    Real-time qRT-PCR analysis of the H3K27 demethylases UTX and JMJD3 , the H3K27 methyltransferase EZH2 and the CDK4/CDK6 inhibitor p16INK4a. Relative mRNA expression levels of UTX , JMJD3 and EZH2 were higher in RCC cancer tissues than in paired adjacent normal tissues (n = 36; all P
    Figure Legend Snippet: Real-time qRT-PCR analysis of the H3K27 demethylases UTX and JMJD3 , the H3K27 methyltransferase EZH2 and the CDK4/CDK6 inhibitor p16INK4a. Relative mRNA expression levels of UTX , JMJD3 and EZH2 were higher in RCC cancer tissues than in paired adjacent normal tissues (n = 36; all P

    Techniques Used: Quantitative RT-PCR, Expressing

    23) Product Images from "Identification of miRNA-mRNA Network Associated with Acute Myeloid Leukemia Survival"

    Article Title: Identification of miRNA-mRNA Network Associated with Acute Myeloid Leukemia Survival

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    doi: 10.12659/MSM.903989

    Verification of expression levels of miRNAs and mRNAs in the low-moderate risk group and the high risk group of AML patients by qRT-PCR. ( A ) IL15RA; ( B ) hsa-miR-200c; ( C ) CD44; ( D ) hsa-miR-425. * is p
    Figure Legend Snippet: Verification of expression levels of miRNAs and mRNAs in the low-moderate risk group and the high risk group of AML patients by qRT-PCR. ( A ) IL15RA; ( B ) hsa-miR-200c; ( C ) CD44; ( D ) hsa-miR-425. * is p

    Techniques Used: Expressing, Quantitative RT-PCR

    24) Product Images from "Long noncoding RNA ILF3-AS1 promotes cell proliferation, migration, and invasion via negatively regulating miR-200b/a/429 in melanoma"

    Article Title: Long noncoding RNA ILF3-AS1 promotes cell proliferation, migration, and invasion via negatively regulating miR-200b/a/429 in melanoma

    Journal: Bioscience Reports

    doi: 10.1042/BSR20171031

    ILF3-AS1 negatively regulates miR-200b/a/429 expression via interacting with EZH2 ( A ) RNA pull-down assays followed by Western blot analysis revealed the specific enrichment of EZH2, but not GAPDH protein with in vitro transcribed biotin-labeled ILF3-AS1 compared with antisense RNA (negative control). ( B ) RIP assays followed by qRT-PCR revealed the specific enrichment of ILF3-AS1, but not GAPDH mRNA with EZH2 antibody compared with nonspecific IgG (negative control). HEIH was used as positive control. ( C ) The specific binding of EZH2 and H3K27me3 levels across the miR-200b/a/429 promoter and the β-actin promoter in ILF3-AS1 stably depleted and control A375 cells were measured by ChIP assays followed by qPCR. ( D ) The expression of miR-200b, miR-200a, and miR-429 in ILF3-AS1 stably depleted and control A375 cells was measured by qRT-PCR. ( E ) The expression of miR-200b, miR-200a, and miR-429 in ILF3-AS1 stably depleted and control SK-MEL-2 cells was measured by qRT-PCR. Data are represented as mean ± SD; * P
    Figure Legend Snippet: ILF3-AS1 negatively regulates miR-200b/a/429 expression via interacting with EZH2 ( A ) RNA pull-down assays followed by Western blot analysis revealed the specific enrichment of EZH2, but not GAPDH protein with in vitro transcribed biotin-labeled ILF3-AS1 compared with antisense RNA (negative control). ( B ) RIP assays followed by qRT-PCR revealed the specific enrichment of ILF3-AS1, but not GAPDH mRNA with EZH2 antibody compared with nonspecific IgG (negative control). HEIH was used as positive control. ( C ) The specific binding of EZH2 and H3K27me3 levels across the miR-200b/a/429 promoter and the β-actin promoter in ILF3-AS1 stably depleted and control A375 cells were measured by ChIP assays followed by qPCR. ( D ) The expression of miR-200b, miR-200a, and miR-429 in ILF3-AS1 stably depleted and control A375 cells was measured by qRT-PCR. ( E ) The expression of miR-200b, miR-200a, and miR-429 in ILF3-AS1 stably depleted and control SK-MEL-2 cells was measured by qRT-PCR. Data are represented as mean ± SD; * P

    Techniques Used: Expressing, Western Blot, In Vitro, Labeling, Negative Control, Quantitative RT-PCR, Positive Control, Binding Assay, Stable Transfection, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction

    Inhibition of miR-200b/a/429 abrogates the inhibitory effects of ILF3-AS1 knockdown on melanoma cell proliferation, migration, and invasion ( A ) The expression of miR-200b, miR-200a, and miR-429 in ILF3-AS1 stably depleted and control A375 cells transfected with miR-200b/a/429 inhibitors or control was measured by qRT-PCR. ( B ) Glo cell viability assays in ILF3-AS1 stably depleted and control A375 cells transfected with miR-200b/a/429 inhibitors or control. ( C ) EdU incorporation assays in ILF3-AS1 stably depleted and control A375 cells transfected with miR-200b/a/429 inhibitors or control. The blue color indicates the nuclei, and the red color indicates EdU-positive nuclei; scale bar = 100 μm. ( D ) Transwell migration assays in ILF3-AS1 stably depleted and control A375 cells transfected with miR-200b/a/429 inhibitors or control. Representative images are shown; scale bar = 100 μm. ( E ) Transwell invasion assays in ILF3-AS1 stably depleted and control A375 cells transfected with miR-200b/a/429 inhibitors or control. Representative images are shown; scale bar = 100 μm. For all panels, data are represented as mean ± SD; ** P
    Figure Legend Snippet: Inhibition of miR-200b/a/429 abrogates the inhibitory effects of ILF3-AS1 knockdown on melanoma cell proliferation, migration, and invasion ( A ) The expression of miR-200b, miR-200a, and miR-429 in ILF3-AS1 stably depleted and control A375 cells transfected with miR-200b/a/429 inhibitors or control was measured by qRT-PCR. ( B ) Glo cell viability assays in ILF3-AS1 stably depleted and control A375 cells transfected with miR-200b/a/429 inhibitors or control. ( C ) EdU incorporation assays in ILF3-AS1 stably depleted and control A375 cells transfected with miR-200b/a/429 inhibitors or control. The blue color indicates the nuclei, and the red color indicates EdU-positive nuclei; scale bar = 100 μm. ( D ) Transwell migration assays in ILF3-AS1 stably depleted and control A375 cells transfected with miR-200b/a/429 inhibitors or control. Representative images are shown; scale bar = 100 μm. ( E ) Transwell invasion assays in ILF3-AS1 stably depleted and control A375 cells transfected with miR-200b/a/429 inhibitors or control. Representative images are shown; scale bar = 100 μm. For all panels, data are represented as mean ± SD; ** P

    Techniques Used: Inhibition, Migration, Expressing, Stable Transfection, Transfection, Quantitative RT-PCR

    ILF3-AS1 is up-regulated in melanoma tissues and cell lines, and indicates poor prognosis of melanoma patients ( A ) MiTranscriptome expression data for ILF3-AS1 across all available NHMEs ( n =4), primary melanomas ( n =33), and metastatic melanomas ( n =228). ( B ) The expression of ILF3-AS1 in 37 benign nevi, 60 primary melanomas, and 25 metastatic melanomas was measured by qRT-PCR. ( C ) The expression of ILF3-AS1 in melanomas categorized based on tumor thickness at diagnosis. For (A)–(C), data are represented as median with interquartile range. P values were acquired by Mann–Whitney U test. ( D ) Kaplan–Meier survival analysis of the correlation between ILF3-AS1 expression and overall survival of melanoma patients. P values were acquired by log-rank test. ( E ) The expression of ILF3-AS1 in human epidermal melanocytes (HEMa-LP) and melanoma cell lines (SK-MEL-2, SK-MEL-28, and A375) was measured by qRT-PCR. Data are represented as mean ± SD; ** P
    Figure Legend Snippet: ILF3-AS1 is up-regulated in melanoma tissues and cell lines, and indicates poor prognosis of melanoma patients ( A ) MiTranscriptome expression data for ILF3-AS1 across all available NHMEs ( n =4), primary melanomas ( n =33), and metastatic melanomas ( n =228). ( B ) The expression of ILF3-AS1 in 37 benign nevi, 60 primary melanomas, and 25 metastatic melanomas was measured by qRT-PCR. ( C ) The expression of ILF3-AS1 in melanomas categorized based on tumor thickness at diagnosis. For (A)–(C), data are represented as median with interquartile range. P values were acquired by Mann–Whitney U test. ( D ) Kaplan–Meier survival analysis of the correlation between ILF3-AS1 expression and overall survival of melanoma patients. P values were acquired by log-rank test. ( E ) The expression of ILF3-AS1 in human epidermal melanocytes (HEMa-LP) and melanoma cell lines (SK-MEL-2, SK-MEL-28, and A375) was measured by qRT-PCR. Data are represented as mean ± SD; ** P

    Techniques Used: Expressing, Quantitative RT-PCR, MANN-WHITNEY

    25) Product Images from "Critical role for the long non-coding RNA AFAP1-AS1 in the proliferation and metastasis of hepatocellular carcinoma"

    Article Title: Critical role for the long non-coding RNA AFAP1-AS1 in the proliferation and metastasis of hepatocellular carcinoma

    Journal: Tumour Biology

    doi: 10.1007/s13277-016-4858-8

    Knockdown of AFAP1-AS1 decreases HCC cell proliferation in vitro. a qRT-PCR was performed to detect the expression of AFAP1-AS1 in MHCC-97 L and MHCC-97H cells transduced with SCR or siAFAP1-AS1. The effects of knockdown of AFAP1-AS1 in cells on cell proliferation b colony formation c and apoptosis d were examined. * P
    Figure Legend Snippet: Knockdown of AFAP1-AS1 decreases HCC cell proliferation in vitro. a qRT-PCR was performed to detect the expression of AFAP1-AS1 in MHCC-97 L and MHCC-97H cells transduced with SCR or siAFAP1-AS1. The effects of knockdown of AFAP1-AS1 in cells on cell proliferation b colony formation c and apoptosis d were examined. * P

    Techniques Used: In Vitro, Quantitative RT-PCR, Expressing, Transduction

    AFAP1-AS1 expression is increased in human HCC tissues and cell lines. a The relative expression of AFAP1-AS1 was detected in 156 pairs of primary HCC tissues and their corresponding adjacent tissues. b qRT-PCR analysis was performed to assess the AFAP1-AS1 levels in HCC cells (SMMC-7721, BEL-7402, MHCC-97 L, and MHCC-97H) and LO2. * P
    Figure Legend Snippet: AFAP1-AS1 expression is increased in human HCC tissues and cell lines. a The relative expression of AFAP1-AS1 was detected in 156 pairs of primary HCC tissues and their corresponding adjacent tissues. b qRT-PCR analysis was performed to assess the AFAP1-AS1 levels in HCC cells (SMMC-7721, BEL-7402, MHCC-97 L, and MHCC-97H) and LO2. * P

    Techniques Used: Expressing, Quantitative RT-PCR

    AFAP1-AS1 silencing inhibited tumor growth in a xenograft mouse model. a Representative photographs of tumors are shown. Tumor weight b and tumor growth curves c in mice are shown for MHCC-97H cells transduced with SCR or siAFAP1-AS1. d qRT-PCR was used to detect the average expression of AFAP1-AS1 in xenograft tumors. e Representative images of IHC staining showed the expression of Ki67 in xenograft tumor tissues from the siAFAP1-AS1 group or SCR group. ** P
    Figure Legend Snippet: AFAP1-AS1 silencing inhibited tumor growth in a xenograft mouse model. a Representative photographs of tumors are shown. Tumor weight b and tumor growth curves c in mice are shown for MHCC-97H cells transduced with SCR or siAFAP1-AS1. d qRT-PCR was used to detect the average expression of AFAP1-AS1 in xenograft tumors. e Representative images of IHC staining showed the expression of Ki67 in xenograft tumor tissues from the siAFAP1-AS1 group or SCR group. ** P

    Techniques Used: Mouse Assay, Transduction, Quantitative RT-PCR, Expressing, Immunohistochemistry, Staining

    26) Product Images from "NF-κB1, c-Rel, and ELK1 inhibit miR-134 expression leading to TAB1 upregulation in paclitaxel-resistant human ovarian cancer"

    Article Title: NF-κB1, c-Rel, and ELK1 inhibit miR-134 expression leading to TAB1 upregulation in paclitaxel-resistant human ovarian cancer

    Journal: Oncotarget

    doi: 10.18632/oncotarget.15267

    NF-κB1, c-Rel and ELK1 bind directly to the response elements in the putative promoter of miR-134 in ovarian cancer cells A . Five genomic regions (R1–R5) spanning a 2.4 kb sequence upstream of the pre-miR-134. B-D . Binding of NF-κB1, c-Rel and ELK1 to the miR-134 promoter region was validated in SKOV3-TR30 cells by ChIP. Non-immune IgG and input DNA served as negative and positive controls, respectively. The enrichment of the binding of NF-κB1, c-Rel and ELK1 with the R3, R5 and R1 regions was quantified from the corresponding ChIP with qPCR. E . EMSA was performed with nuclear extracts from SKOV3-TR30 cells incubated with 5′-biotin-labeled oligonucleotide sequences containing the binding sites for the NF-κB1, c-Rel and ELK1 transcription factors. Unlabeled competitor sequence was also included to indicate the specificity of the protein-DNA complexes. F . pBI- NF-κB1, pBI- c-Rel and pBI- ELK1 overexpression plasmids were transfected into SKOV3 cells; the transfection efficiency was validated by qRT-PCR and Western blot analyses. G . At 48 h after transfection, luciferase activity was determined and then normalized to Renilla values. H . Sequencing was used to identify that the plasmids including pGL3-promoter-R1 (containing the ELK1 binding site) and its mutant pGL3-promoter-R1-mut, pGL3-promoter-R3 (containing the NF-κB1 binding site) and its mutant pGL3-promoter-R3-mut as well as pGL3-promoter-R5 (containing the c-Rel binding site) and its mutant pGL3-promoter-R5-mut were successfully constructed for luciferase reporter assay. Data represent the mean ± SE of three independent experiments(* P
    Figure Legend Snippet: NF-κB1, c-Rel and ELK1 bind directly to the response elements in the putative promoter of miR-134 in ovarian cancer cells A . Five genomic regions (R1–R5) spanning a 2.4 kb sequence upstream of the pre-miR-134. B-D . Binding of NF-κB1, c-Rel and ELK1 to the miR-134 promoter region was validated in SKOV3-TR30 cells by ChIP. Non-immune IgG and input DNA served as negative and positive controls, respectively. The enrichment of the binding of NF-κB1, c-Rel and ELK1 with the R3, R5 and R1 regions was quantified from the corresponding ChIP with qPCR. E . EMSA was performed with nuclear extracts from SKOV3-TR30 cells incubated with 5′-biotin-labeled oligonucleotide sequences containing the binding sites for the NF-κB1, c-Rel and ELK1 transcription factors. Unlabeled competitor sequence was also included to indicate the specificity of the protein-DNA complexes. F . pBI- NF-κB1, pBI- c-Rel and pBI- ELK1 overexpression plasmids were transfected into SKOV3 cells; the transfection efficiency was validated by qRT-PCR and Western blot analyses. G . At 48 h after transfection, luciferase activity was determined and then normalized to Renilla values. H . Sequencing was used to identify that the plasmids including pGL3-promoter-R1 (containing the ELK1 binding site) and its mutant pGL3-promoter-R1-mut, pGL3-promoter-R3 (containing the NF-κB1 binding site) and its mutant pGL3-promoter-R3-mut as well as pGL3-promoter-R5 (containing the c-Rel binding site) and its mutant pGL3-promoter-R5-mut were successfully constructed for luciferase reporter assay. Data represent the mean ± SE of three independent experiments(* P

    Techniques Used: Sequencing, Binding Assay, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Incubation, Labeling, Over Expression, Transfection, Quantitative RT-PCR, Western Blot, Luciferase, Activity Assay, Mutagenesis, Construct, Reporter Assay

    Correlations between miR-134 levels and expression of NF-κB1, c-Rel and ELK1 in serous EOC specimens A . NF-κB1, c-Rel and ELK1 mRNA expression was analyzed by qRT-PCR in tissue from 24 cases of chemosensitive serous EOC and from 24 cases of chemoresistant serous EOC. NF-κB1 mRNA expression was significantly upregulated in chemoresistant serous EOC tissues, while c-Rel mRNA and ELK1 mRNA were overexpressed in chemoresistant tissues compared with that in chemosensitive tissues. B . Spearman correlation coefficient analyses of the correlations between miR-134 and expression of NF-κB1, c-Rel and ELK1 were performed for data obtained from qRT-PCR results and the correlation coefficient ‘r’ was calculated. C-D . Immunohistochemical staining of NF-κB1, c-Rel and ELK1 in serous epithelial ovarian cancer tissues. C, Expression of NF-κB1, c-Rel and ELK1 in chemoresistant tissue. D, Expression of NF-κB1, c-Rel and ELK1 in chemosensitive tissue. E . Patients with high nucleus NF-κB1, c-Rel and ELK1 expression showed significantly shorter overall survival than those with low nucleus expression. F . The positive cytoplasmic expression of NF-κB1, c-Rel and ELK1 showed no effect on survival in serous EOC patients.
    Figure Legend Snippet: Correlations between miR-134 levels and expression of NF-κB1, c-Rel and ELK1 in serous EOC specimens A . NF-κB1, c-Rel and ELK1 mRNA expression was analyzed by qRT-PCR in tissue from 24 cases of chemosensitive serous EOC and from 24 cases of chemoresistant serous EOC. NF-κB1 mRNA expression was significantly upregulated in chemoresistant serous EOC tissues, while c-Rel mRNA and ELK1 mRNA were overexpressed in chemoresistant tissues compared with that in chemosensitive tissues. B . Spearman correlation coefficient analyses of the correlations between miR-134 and expression of NF-κB1, c-Rel and ELK1 were performed for data obtained from qRT-PCR results and the correlation coefficient ‘r’ was calculated. C-D . Immunohistochemical staining of NF-κB1, c-Rel and ELK1 in serous epithelial ovarian cancer tissues. C, Expression of NF-κB1, c-Rel and ELK1 in chemoresistant tissue. D, Expression of NF-κB1, c-Rel and ELK1 in chemosensitive tissue. E . Patients with high nucleus NF-κB1, c-Rel and ELK1 expression showed significantly shorter overall survival than those with low nucleus expression. F . The positive cytoplasmic expression of NF-κB1, c-Rel and ELK1 showed no effect on survival in serous EOC patients.

    Techniques Used: Expressing, Quantitative RT-PCR, Immunohistochemistry, Staining

    27) Product Images from "Circular RNA circ-ITCH inhibits bladder cancer progression by sponging miR-17/miR-224 and regulating p21, PTEN expression"

    Article Title: Circular RNA circ-ITCH inhibits bladder cancer progression by sponging miR-17/miR-224 and regulating p21, PTEN expression

    Journal: Molecular Cancer

    doi: 10.1186/s12943-018-0771-7

    miR-17 and miR-224 played as oncogenes in BCa and eliminated the repression function of cir-ITCH. a and b miR-17 and miR-224 were up-regulated in BCa tissues as compared with adjacent normal tissues using qRT-PCR ( n = 28, ** P
    Figure Legend Snippet: miR-17 and miR-224 played as oncogenes in BCa and eliminated the repression function of cir-ITCH. a and b miR-17 and miR-224 were up-regulated in BCa tissues as compared with adjacent normal tissues using qRT-PCR ( n = 28, ** P

    Techniques Used: BIA-KA, Quantitative RT-PCR

    Cir-ITCH modulated the expression of endogenous miR-17 and miR-224 targets p21 and PTEN. a Putative miR-17/miR-224 binding sequence in the 3′-UTR of p21 and PTEN mRNA. b and c cir-ITCH up-regulated the mRNA expression level of p21 and PTEN in BCa cells by qRT-PCR (* P
    Figure Legend Snippet: Cir-ITCH modulated the expression of endogenous miR-17 and miR-224 targets p21 and PTEN. a Putative miR-17/miR-224 binding sequence in the 3′-UTR of p21 and PTEN mRNA. b and c cir-ITCH up-regulated the mRNA expression level of p21 and PTEN in BCa cells by qRT-PCR (* P

    Techniques Used: Expressing, Binding Assay, Sequencing, BIA-KA, Quantitative RT-PCR

    Cir-ITCH was significantly decreased in BCa and correlated with prognosis of BCa patients. a and b Relative expression level of cir-ITCH in BCa tissues (T) and adjacent normal tissues (N) (a: n = 72, b: n = 59) using qRT-PCR. cir-ITCH expression was significantly lower in bladder cancer tissues, compared with that in adjacent normal tissues (* P
    Figure Legend Snippet: Cir-ITCH was significantly decreased in BCa and correlated with prognosis of BCa patients. a and b Relative expression level of cir-ITCH in BCa tissues (T) and adjacent normal tissues (N) (a: n = 72, b: n = 59) using qRT-PCR. cir-ITCH expression was significantly lower in bladder cancer tissues, compared with that in adjacent normal tissues (* P

    Techniques Used: BIA-KA, Expressing, Quantitative RT-PCR

    Cir-ITCH acted as a sponge for miR-17 and miR-224. a Cir-ITCH in BCa cell lysis was pulled down and enriched with cir-ITCH specific probe and then detected by qRT-PCR(** P
    Figure Legend Snippet: Cir-ITCH acted as a sponge for miR-17 and miR-224. a Cir-ITCH in BCa cell lysis was pulled down and enriched with cir-ITCH specific probe and then detected by qRT-PCR(** P

    Techniques Used: BIA-KA, Lysis, Quantitative RT-PCR

    28) Product Images from "miR-20a regulates proliferation, differentiation and apoptosis in P19 cell model of cardiac differentiation by targeting Smoothened"

    Article Title: miR-20a regulates proliferation, differentiation and apoptosis in P19 cell model of cardiac differentiation by targeting Smoothened

    Journal: Biology Open

    doi: 10.1242/bio.019182

    miR-20a is downregulated and SMO is upregulated in myocardially differentiated P19 cells. (A) Western blot assay shows that the expression levels of marker proteins of cardiomyocytes cTnT, GATA4 and desmin were obviously higher in P19 cells at differentiation day 10 than those in P19 cells at differentiation day 0. (B) qRT-PCR shows that compared with P19 cells at differentiation day 0, P19 cells at differentiation day 10 have an obvious decrease in miR-20a expression. (C) The expression level of SMO protein is significantly higher in P19 cells at differentiation day 10 than that in P19 cells at differentiation day 0. Error bars indicate s.d., n =3. * P
    Figure Legend Snippet: miR-20a is downregulated and SMO is upregulated in myocardially differentiated P19 cells. (A) Western blot assay shows that the expression levels of marker proteins of cardiomyocytes cTnT, GATA4 and desmin were obviously higher in P19 cells at differentiation day 10 than those in P19 cells at differentiation day 0. (B) qRT-PCR shows that compared with P19 cells at differentiation day 0, P19 cells at differentiation day 10 have an obvious decrease in miR-20a expression. (C) The expression level of SMO protein is significantly higher in P19 cells at differentiation day 10 than that in P19 cells at differentiation day 0. Error bars indicate s.d., n =3. * P

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

    29) Product Images from "Epstein-Barr virus encoded miR-BART11 promotes inflammation-induced carcinogenesis by targeting FOXP1"

    Article Title: Epstein-Barr virus encoded miR-BART11 promotes inflammation-induced carcinogenesis by targeting FOXP1

    Journal: Oncotarget

    doi: 10.18632/oncotarget.9170

    EBV-miR-BART11 promotes monocyte differentiation of THP-1 cells by attenuating FOXP1 expression A. FOXP1 expression at mRNA (left) and protein (right) levels, during the PMA-induced differentiation of monocytic THP-1 cells. B. The expression of EBV-miR-BART11 (left) and FOXP1 (middle, mRNA; right, protein) was examined by qRT-PCR and western blotting, respectively, in THP-1 cells infected with lentivirus encoding EBV-miR-BART11. C. The effects of EBV-miR-BART11 and FOXP1 on monocyte differentiation. Morphological changes were monitored in PMA-induced THP-1 cells following FOXP1 overexpression vector or EBV-miR-BART11 precursor vector transfection. THP-1 cell differentiation was determined by the viability of adherent cells, using MTT assay. Data represent mean ± SD of OD values obtained in three separate experiments (ND: not detected; ** p
    Figure Legend Snippet: EBV-miR-BART11 promotes monocyte differentiation of THP-1 cells by attenuating FOXP1 expression A. FOXP1 expression at mRNA (left) and protein (right) levels, during the PMA-induced differentiation of monocytic THP-1 cells. B. The expression of EBV-miR-BART11 (left) and FOXP1 (middle, mRNA; right, protein) was examined by qRT-PCR and western blotting, respectively, in THP-1 cells infected with lentivirus encoding EBV-miR-BART11. C. The effects of EBV-miR-BART11 and FOXP1 on monocyte differentiation. Morphological changes were monitored in PMA-induced THP-1 cells following FOXP1 overexpression vector or EBV-miR-BART11 precursor vector transfection. THP-1 cell differentiation was determined by the viability of adherent cells, using MTT assay. Data represent mean ± SD of OD values obtained in three separate experiments (ND: not detected; ** p

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Infection, Over Expression, Plasmid Preparation, Transfection, Cell Differentiation, MTT Assay

    FOXP1 is a direct target of EBV-miR-BART11 A. Three binding sites of EBV-miR-BART11-3p and EBV-miR-BART11-5p were predicted in the FOXP1 3′-UTR, including 3082 bp-3106 bp (I), 3917 bp-3998 bp (II), and 5943 bp-5970 bp (III). Wild-type (FOXP1-WT) and mutant (FOXP1-mutant) sequences were used to validate these predictions. B. The expression of exogenous EBV-miR-BART11-3p (left) and EBV-miR-BART11-5p (right) was detected by qRT-PCR. FOXP1 C. mRNA and D. protein expression levels in 5-8F, HK-1, and AGS cells after EBV-miR-BART11 treatment. β-actin served as loading control. E. Luciferase reporter assay, using reporter vectors containing either wild-type (FOXP1-WT) or mutant (FOXP1-mutant) FOXP1 3′-UTR, and EBV-miR-BART11 or non-targeting control, was performed in order to identify the direct binding of EBV-miR-BART11 to the FOXP1 3′-UTR in 5-8F cells. F. EBV-miR-BART11 and FOXP1 expression levels in NPC and control specimens were detected by qRT-PCR. N, non-tumor nasopharyngeal epithelium (n = 10); T, NPC (n = 30). Representative images or data expressed as mean ± SD of the measurements obtained in three separate experiments are presented (ND: not detected; * p
    Figure Legend Snippet: FOXP1 is a direct target of EBV-miR-BART11 A. Three binding sites of EBV-miR-BART11-3p and EBV-miR-BART11-5p were predicted in the FOXP1 3′-UTR, including 3082 bp-3106 bp (I), 3917 bp-3998 bp (II), and 5943 bp-5970 bp (III). Wild-type (FOXP1-WT) and mutant (FOXP1-mutant) sequences were used to validate these predictions. B. The expression of exogenous EBV-miR-BART11-3p (left) and EBV-miR-BART11-5p (right) was detected by qRT-PCR. FOXP1 C. mRNA and D. protein expression levels in 5-8F, HK-1, and AGS cells after EBV-miR-BART11 treatment. β-actin served as loading control. E. Luciferase reporter assay, using reporter vectors containing either wild-type (FOXP1-WT) or mutant (FOXP1-mutant) FOXP1 3′-UTR, and EBV-miR-BART11 or non-targeting control, was performed in order to identify the direct binding of EBV-miR-BART11 to the FOXP1 3′-UTR in 5-8F cells. F. EBV-miR-BART11 and FOXP1 expression levels in NPC and control specimens were detected by qRT-PCR. N, non-tumor nasopharyngeal epithelium (n = 10); T, NPC (n = 30). Representative images or data expressed as mean ± SD of the measurements obtained in three separate experiments are presented (ND: not detected; * p

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

    30) Product Images from "Mitochondrial DNA 10609T Promotes Hypoxia-Induced Increase of Intracellular ROS and Is a Risk Factor of High Altitude Polycythemia"

    Article Title: Mitochondrial DNA 10609T Promotes Hypoxia-Induced Increase of Intracellular ROS and Is a Risk Factor of High Altitude Polycythemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0087775

    The mRNA level of EPO in cybrids. A. qRT-PCR analysis of mRNA level of EPO from 143B-10609T (WT) and 143B-10609C (MU) cultured for 12 h under hypoxic conditions (37°C, 5% carbon dioxide, and 3% oxygen) (n = 3). B. qRT-PCR analysis of mRNA level of EPO from 143B-10609T (WT) incubated in the absence or presence of 2.5 mM N-acetyl-L-cysteine (NAC) for 12 h under hypoxic conditions (37°C, 5% carbon dioxide, and 3% oxygen) (n = 3). Housekeeping gene was beta-actin, and the relative expression value was calculated by 2−ΔΔT method. Error bars indicate the standard deviation. * P
    Figure Legend Snippet: The mRNA level of EPO in cybrids. A. qRT-PCR analysis of mRNA level of EPO from 143B-10609T (WT) and 143B-10609C (MU) cultured for 12 h under hypoxic conditions (37°C, 5% carbon dioxide, and 3% oxygen) (n = 3). B. qRT-PCR analysis of mRNA level of EPO from 143B-10609T (WT) incubated in the absence or presence of 2.5 mM N-acetyl-L-cysteine (NAC) for 12 h under hypoxic conditions (37°C, 5% carbon dioxide, and 3% oxygen) (n = 3). Housekeeping gene was beta-actin, and the relative expression value was calculated by 2−ΔΔT method. Error bars indicate the standard deviation. * P

    Techniques Used: Quantitative RT-PCR, Cell Culture, Incubation, Expressing, Standard Deviation

    31) Product Images from "Targeting Wnt pathway in mantle cell lymphoma-initiating cells"

    Article Title: Targeting Wnt pathway in mantle cell lymphoma-initiating cells

    Journal: Journal of Hematology & Oncology

    doi: 10.1186/s13045-015-0161-1

    Stem cell-like properties of MCL-ICs. a – c qRT-PCR performed using the total cellular RNA isolated from MCL-ICs ( n = 4) for a stem cell transcription factors (Nanog, Oct4, Sox2, Klf4), b ALDH isoforms and antioxidant enzymes SOD2 and MT1b, and c chemoresistance-associated genes encoding ABCC3, ABCC6, and CD44. Differences between MCL-ICs and MCL-non-ICs were significant ( P
    Figure Legend Snippet: Stem cell-like properties of MCL-ICs. a – c qRT-PCR performed using the total cellular RNA isolated from MCL-ICs ( n = 4) for a stem cell transcription factors (Nanog, Oct4, Sox2, Klf4), b ALDH isoforms and antioxidant enzymes SOD2 and MT1b, and c chemoresistance-associated genes encoding ABCC3, ABCC6, and CD44. Differences between MCL-ICs and MCL-non-ICs were significant ( P

    Techniques Used: Quantitative RT-PCR, Isolation

    Isolation of MCL-ICs. ( a ) Isolation of MCL-ICs using immunostaining and flow sorting. ( b ) Immunostaining of isolated MCL-ICs for plasma cell markers CD27/CD38 and natural killer cell markers CD56/CD16 detected by flow cytometry. ( c ) Detection of gene fusion t (11;14) (q13; q32) in MCL-ICs using fluorescent in situ hybridization, indicated by arrow . ( d ) qRT-PCR expression of cyclin D1 in MCL-ICs, MCL-non-ICs relative to B-cells. Differences between MCL-ICs and B-cells were significant ( P
    Figure Legend Snippet: Isolation of MCL-ICs. ( a ) Isolation of MCL-ICs using immunostaining and flow sorting. ( b ) Immunostaining of isolated MCL-ICs for plasma cell markers CD27/CD38 and natural killer cell markers CD56/CD16 detected by flow cytometry. ( c ) Detection of gene fusion t (11;14) (q13; q32) in MCL-ICs using fluorescent in situ hybridization, indicated by arrow . ( d ) qRT-PCR expression of cyclin D1 in MCL-ICs, MCL-non-ICs relative to B-cells. Differences between MCL-ICs and B-cells were significant ( P

    Techniques Used: Isolation, Immunostaining, Flow Cytometry, Cytometry, In Situ Hybridization, Quantitative RT-PCR, Expressing

    32) Product Images from "CLDN10 is Associated with Papillary Thyroid Cancer Progression"

    Article Title: CLDN10 is Associated with Papillary Thyroid Cancer Progression

    Journal: Journal of Cancer

    doi: 10.7150/jca.28636

    CLDN10 is up-regulated in human PTC tissues and cell lines. (a) Relationship of expression levels of CLDN10 in thyroid tumor tissues and normal tissues in TCGA. (b) CLDN10 is significantly increased in 55 human PTC tissues in comparison to matched adjacent tissues. (c) The relative expression of CLDN10 to GAPDH using qRT-PCR. Both B-CPAP and KTC-1 cell lines are overexpressed. (d) The efficiency of siRNAs (Si-NC, Si-CLDN10#1 and Si-CLDN10#2) was assayed by qRT-PCR in B-CPAPA and KTC-1 cells. *P
    Figure Legend Snippet: CLDN10 is up-regulated in human PTC tissues and cell lines. (a) Relationship of expression levels of CLDN10 in thyroid tumor tissues and normal tissues in TCGA. (b) CLDN10 is significantly increased in 55 human PTC tissues in comparison to matched adjacent tissues. (c) The relative expression of CLDN10 to GAPDH using qRT-PCR. Both B-CPAP and KTC-1 cell lines are overexpressed. (d) The efficiency of siRNAs (Si-NC, Si-CLDN10#1 and Si-CLDN10#2) was assayed by qRT-PCR in B-CPAPA and KTC-1 cells. *P

    Techniques Used: Expressing, Quantitative RT-PCR

    33) Product Images from "Effects of Antipsychotic Drugs on the Epigenetic Modification of Brain-Derived Neurotrophic Factor Gene Expression in the Hippocampi of Chronic Restraint Stress Rats"

    Article Title: Effects of Antipsychotic Drugs on the Epigenetic Modification of Brain-Derived Neurotrophic Factor Gene Expression in the Hippocampi of Chronic Restraint Stress Rats

    Journal: Neural Plasticity

    doi: 10.1155/2018/2682037

    Effects of antipsychotic drugs on histone deacetylase 5 (HDAC5) mRNA levels in the rat hippocampus. Rats were given a daily injection of either VEH, OLA, or HAL for 21 d in conjunction with either + stress or − stress. HDAC5 mRNA levels in the rat hippocampus were assessed using a qRT-PCR procedure. The quantitative analysis was normalized to GAPDH. ∗∗ p
    Figure Legend Snippet: Effects of antipsychotic drugs on histone deacetylase 5 (HDAC5) mRNA levels in the rat hippocampus. Rats were given a daily injection of either VEH, OLA, or HAL for 21 d in conjunction with either + stress or − stress. HDAC5 mRNA levels in the rat hippocampus were assessed using a qRT-PCR procedure. The quantitative analysis was normalized to GAPDH. ∗∗ p

    Techniques Used: Histone Deacetylase Assay, Injection, Quantitative RT-PCR

    Effects of antipsychotic drugs on DNA methyltransferase (DNMT) 1 and DNMT3a mRNA levels in the rat hippocampus. Rats were given a daily injection of either VEH, OLA, or HAL for 21 d in conjunction with either + stress or − stress. The DNMT1 (a) and DNMT3a (b) mRNA levels in the rat hippocampus were assessed using a qRT-PCR procedure. The quantitative analysis was normalized to GAPDH. ∗ p
    Figure Legend Snippet: Effects of antipsychotic drugs on DNA methyltransferase (DNMT) 1 and DNMT3a mRNA levels in the rat hippocampus. Rats were given a daily injection of either VEH, OLA, or HAL for 21 d in conjunction with either + stress or − stress. The DNMT1 (a) and DNMT3a (b) mRNA levels in the rat hippocampus were assessed using a qRT-PCR procedure. The quantitative analysis was normalized to GAPDH. ∗ p

    Techniques Used: Injection, Quantitative RT-PCR

    Effects of antipsychotic drugs on acetylated histone H3 and methyl CpG-binding protein 2 (MeCP2) levels at BDNF promoter IV in the rat hippocampus. Rats were given a daily injection of either VEH, OLA, or HAL for 21 d in conjunction with either + stress or − stress. Chromatin immunoprecipitation (ChIP) assays were performed to measure the levels of acetylated H3 (a) and MeCP2 (b) at BDNF promoter IV in the rat hippocampus using specific antibodies. These levels were quantified by a qRT-PCR procedure. ∗∗ p
    Figure Legend Snippet: Effects of antipsychotic drugs on acetylated histone H3 and methyl CpG-binding protein 2 (MeCP2) levels at BDNF promoter IV in the rat hippocampus. Rats were given a daily injection of either VEH, OLA, or HAL for 21 d in conjunction with either + stress or − stress. Chromatin immunoprecipitation (ChIP) assays were performed to measure the levels of acetylated H3 (a) and MeCP2 (b) at BDNF promoter IV in the rat hippocampus using specific antibodies. These levels were quantified by a qRT-PCR procedure. ∗∗ p

    Techniques Used: Binding Assay, Injection, Chromatin Immunoprecipitation, Quantitative RT-PCR

    34) Product Images from "Overexpression of HDAC9 is associated with poor prognosis and tumor progression of breast cancer in Chinese females"

    Article Title: Overexpression of HDAC9 is associated with poor prognosis and tumor progression of breast cancer in Chinese females

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S164583

    The mRNA expression of HDAC9 measured by qRT-PCR. Notes: ( A ) HDAC9 expression in breast cancer tissues and paired normal tissues. The expression of HDAC9 was higher in breast cancer tissues than that in the matched normal tissues (*** P
    Figure Legend Snippet: The mRNA expression of HDAC9 measured by qRT-PCR. Notes: ( A ) HDAC9 expression in breast cancer tissues and paired normal tissues. The expression of HDAC9 was higher in breast cancer tissues than that in the matched normal tissues (*** P

    Techniques Used: Expressing, Quantitative RT-PCR

    35) Product Images from "Long non-coding RNA HOTAIR promotes UVB-induced apoptosis and inflammatory injury by up-regulation of PKR in keratinocytes"

    Article Title: Long non-coding RNA HOTAIR promotes UVB-induced apoptosis and inflammatory injury by up-regulation of PKR in keratinocytes

    Journal: Brazilian Journal of Medical and Biological Research

    doi: 10.1590/1414-431X20186896

    Effect of HOTAIR on UVB-induced cell injury; A , qRT-PCR revealed overexpression and suppression of HOTAIR in HaCaT cells after transfection assay; B , Cell viability assay revealed suppression of HOTAIR alleviated UVB-induced low cell viability and overexpression of HOTAIR further promoted UVB-induced inhibition of cell viability; C , Apoptosis assay revealed suppression of HOTAIR alleviated UVB-induced high cell apoptosis and overexpression of HOTAIR further promoted UVB-induced high cell apoptosis. D , qRT-PCR analysis showed that mRNA expression of Bcl-2 was increased after HOTAIR was silenced and decreased after HOTAIR was overexpressed. E , Western blotting analysis showed increased expression of Bax and decreased expression of Bcl-2 in HOTAIR overexpressed cells; F , ELISA results showed suppression of HOTAIR alleviated UVB-induced high expression of TNF-α and overexpression of HOTAIR further promoted UVB-induced high expression of TNF-α; G , ELISA results showed suppression of HOTAIR alleviated UVB-induced high expression of IL-6 and overexpression of HOTAIR further promoted UVB-induced high expression of IL-6; H , Western blotting analysis showed increased expression of TNF-α and IL-6 with HOTAIR overexpression. Data are reported as means±SD. *P
    Figure Legend Snippet: Effect of HOTAIR on UVB-induced cell injury; A , qRT-PCR revealed overexpression and suppression of HOTAIR in HaCaT cells after transfection assay; B , Cell viability assay revealed suppression of HOTAIR alleviated UVB-induced low cell viability and overexpression of HOTAIR further promoted UVB-induced inhibition of cell viability; C , Apoptosis assay revealed suppression of HOTAIR alleviated UVB-induced high cell apoptosis and overexpression of HOTAIR further promoted UVB-induced high cell apoptosis. D , qRT-PCR analysis showed that mRNA expression of Bcl-2 was increased after HOTAIR was silenced and decreased after HOTAIR was overexpressed. E , Western blotting analysis showed increased expression of Bax and decreased expression of Bcl-2 in HOTAIR overexpressed cells; F , ELISA results showed suppression of HOTAIR alleviated UVB-induced high expression of TNF-α and overexpression of HOTAIR further promoted UVB-induced high expression of TNF-α; G , ELISA results showed suppression of HOTAIR alleviated UVB-induced high expression of IL-6 and overexpression of HOTAIR further promoted UVB-induced high expression of IL-6; H , Western blotting analysis showed increased expression of TNF-α and IL-6 with HOTAIR overexpression. Data are reported as means±SD. *P

    Techniques Used: Quantitative RT-PCR, Over Expression, Transfection, Viability Assay, Inhibition, Apoptosis Assay, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

    Effect of PKR on UVB-induced cell injury; A qRT-PCR revealed downregulation of PKR in HaCaT cells; B, Western blotting analysis revealed downregulation of PKR in HaCaT cells; C , qRT-PCR revealed overexpression of PKR in HaCaT cells; D , Western blotting analysis revealed overexpression of PKR in HaCaT cells; E , Cell viability assay revealed overexpression of PKR promoted UVB-induced cell viability inhibition, and the effects were reduced by suppression of PKR; F , Apoptosis assay revealed overexpression of PKR promoted UVB-induced cell apoptosis, and the effects were reduced by suppression of PKR; G , Western blotting analysis revealed overexpression of PKR promoted UVB-induced alterations in apoptosis-associated factors, and the effects were reduced by suppression of PKR. Data are reported as means±SD. *P
    Figure Legend Snippet: Effect of PKR on UVB-induced cell injury; A qRT-PCR revealed downregulation of PKR in HaCaT cells; B, Western blotting analysis revealed downregulation of PKR in HaCaT cells; C , qRT-PCR revealed overexpression of PKR in HaCaT cells; D , Western blotting analysis revealed overexpression of PKR in HaCaT cells; E , Cell viability assay revealed overexpression of PKR promoted UVB-induced cell viability inhibition, and the effects were reduced by suppression of PKR; F , Apoptosis assay revealed overexpression of PKR promoted UVB-induced cell apoptosis, and the effects were reduced by suppression of PKR; G , Western blotting analysis revealed overexpression of PKR promoted UVB-induced alterations in apoptosis-associated factors, and the effects were reduced by suppression of PKR. Data are reported as means±SD. *P

    Techniques Used: Quantitative RT-PCR, Western Blot, Over Expression, Viability Assay, Inhibition, Apoptosis Assay

    qRT-PCR was performed to evaluate the effect of UVB on HOTAIR and data revealed that UVB upregulated the expression of HOTAIR. Data are reported as means±SD. ***P
    Figure Legend Snippet: qRT-PCR was performed to evaluate the effect of UVB on HOTAIR and data revealed that UVB upregulated the expression of HOTAIR. Data are reported as means±SD. ***P

    Techniques Used: Quantitative RT-PCR, Expressing

    Effect of HOTAIR on PKR expression; A , qRT-PCR revealed overexpression of HOTAIR promoted the expression of PKR; B , Western blotting analysis revealed overexpression of HOTAIR promoted the expression of PKR. Data are reported as means±SD. *P
    Figure Legend Snippet: Effect of HOTAIR on PKR expression; A , qRT-PCR revealed overexpression of HOTAIR promoted the expression of PKR; B , Western blotting analysis revealed overexpression of HOTAIR promoted the expression of PKR. Data are reported as means±SD. *P

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

    36) Product Images from "Activation of AMPK inhibits TGF-β1-induced airway smooth muscle cells proliferation and its potential mechanisms"

    Article Title: Activation of AMPK inhibits TGF-β1-induced airway smooth muscle cells proliferation and its potential mechanisms

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-21812-0

    Smad2/3 mediates TGF-β1-induced alterations of miR-206, HDAC4 and cyclin D1. ASMCs were treated with SB431542 (10 μM) for 1 h before stimulation with TGF-β1 (10 ng/ml) for 24 h. ( a ) The level of miR-206 was measured using qRT-PCR. U6 small nuclear RNA served as a loading control (n = 4 per group). ( b ) HDAC4 protein level was analyzed using immunoblotting (n = 4 per group). ( c ) Cyclin D1 protein level was determined using immunoblotting (n = 4 per group). The full-length blots of Fig. 2b and 2c are presented in Supplementary Fig. S1. *P
    Figure Legend Snippet: Smad2/3 mediates TGF-β1-induced alterations of miR-206, HDAC4 and cyclin D1. ASMCs were treated with SB431542 (10 μM) for 1 h before stimulation with TGF-β1 (10 ng/ml) for 24 h. ( a ) The level of miR-206 was measured using qRT-PCR. U6 small nuclear RNA served as a loading control (n = 4 per group). ( b ) HDAC4 protein level was analyzed using immunoblotting (n = 4 per group). ( c ) Cyclin D1 protein level was determined using immunoblotting (n = 4 per group). The full-length blots of Fig. 2b and 2c are presented in Supplementary Fig. S1. *P

    Techniques Used: Quantitative RT-PCR

    miR-206 regulates HDAC4/cyclin D1 expression in ASMCs. ( a ) ASMCs were transfected with miR-206 mimics or miR-NC for 48 h, the expression of miR-206 was examined by qRT-PCR. U6 small nuclear RNA served as a loading control (n = 4 per group). ( b ) ASMCs were transfected with miR-206 inhibitors or miR-NC for 48 h, the level of miR-206 was examined by qRT-PCR. U6 small nuclear RNA served as a loading control (n = 4 per group). ASMCs were transfected with miR-206 mimics, miR-206 inhibitors or miR-NC for 48 h. ( c ) HDAC4 protein level were analyzed using immunoblotting (n = 4 per group). ( d ) Cyclin D1 protein level was determined using immunoblotting (n = 4 per group). ( e ) ASMCs were transfected with sequence-specific HDAC4 siRNA or non-targeting siRNA for 48 h, HDAC4 protein level was examined using immunoblotting (n = 4 per group). ( f ) ASMCs were transfected with sequence-specific HDAC4 siRNA or non-targeting siRNA for 48 h, cyclin D1 protein level was determined by immunoblotting (n = 4 per group). The full-length blots of Fig. 3c–f are presented in Supplementary Fig. S2. *P
    Figure Legend Snippet: miR-206 regulates HDAC4/cyclin D1 expression in ASMCs. ( a ) ASMCs were transfected with miR-206 mimics or miR-NC for 48 h, the expression of miR-206 was examined by qRT-PCR. U6 small nuclear RNA served as a loading control (n = 4 per group). ( b ) ASMCs were transfected with miR-206 inhibitors or miR-NC for 48 h, the level of miR-206 was examined by qRT-PCR. U6 small nuclear RNA served as a loading control (n = 4 per group). ASMCs were transfected with miR-206 mimics, miR-206 inhibitors or miR-NC for 48 h. ( c ) HDAC4 protein level were analyzed using immunoblotting (n = 4 per group). ( d ) Cyclin D1 protein level was determined using immunoblotting (n = 4 per group). ( e ) ASMCs were transfected with sequence-specific HDAC4 siRNA or non-targeting siRNA for 48 h, HDAC4 protein level was examined using immunoblotting (n = 4 per group). ( f ) ASMCs were transfected with sequence-specific HDAC4 siRNA or non-targeting siRNA for 48 h, cyclin D1 protein level was determined by immunoblotting (n = 4 per group). The full-length blots of Fig. 3c–f are presented in Supplementary Fig. S2. *P

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

    The mechanisms underlying activation of AMPK inhibition of TGF-β1-induced ASMCs proliferation. ( a ) ASMCs were transfected with AMPK α2-specific or non-targeting siRNA for 48 h, and then treated with metformin (10 mM) for 6 h before stimulation with TGF-β1 (10 ng/ml) for 1 h. The phosphorylation of Smad2/3 was determined by immunoblotting (n = 4 per group). ASMCs were transfected with AMPK α2-specific or non-targeting siRNA for 24 h, and then treated with metformin (10 mM) for 6 h before stimulation with TGF-β1 (10 ng/ml) for 24 h. ( b ) The expression of miR-206 was examined by qRT-PCR (n = 4 per group). ( c ) The expression and phosphorylation of HDAC4 were analyzed using immunoblotting (n = 4 per group). ( d ) Cyclin D1 protein level was determined using immunoblotting (n = 4 per group). The full-length blots of Fig. 6a, 6c and 6d are presented in Supplementary Fig. S4. *P
    Figure Legend Snippet: The mechanisms underlying activation of AMPK inhibition of TGF-β1-induced ASMCs proliferation. ( a ) ASMCs were transfected with AMPK α2-specific or non-targeting siRNA for 48 h, and then treated with metformin (10 mM) for 6 h before stimulation with TGF-β1 (10 ng/ml) for 1 h. The phosphorylation of Smad2/3 was determined by immunoblotting (n = 4 per group). ASMCs were transfected with AMPK α2-specific or non-targeting siRNA for 24 h, and then treated with metformin (10 mM) for 6 h before stimulation with TGF-β1 (10 ng/ml) for 24 h. ( b ) The expression of miR-206 was examined by qRT-PCR (n = 4 per group). ( c ) The expression and phosphorylation of HDAC4 were analyzed using immunoblotting (n = 4 per group). ( d ) Cyclin D1 protein level was determined using immunoblotting (n = 4 per group). The full-length blots of Fig. 6a, 6c and 6d are presented in Supplementary Fig. S4. *P

    Techniques Used: Activation Assay, Inhibition, Transfection, Expressing, Quantitative RT-PCR

    37) Product Images from "Genomic Analysis of Invasive Human Bone Marrow Derived Mesenchymal Stem Cells"

    Article Title: Genomic Analysis of Invasive Human Bone Marrow Derived Mesenchymal Stem Cells

    Journal: Journal of bone marrow research

    doi: 10.4172/2329-8820.1000122

    Gene expression changes in invasive hMSCs A) Microscopic images (20X) of cells invaded toward SCM after 24 hours using either control inserts or inserts containing Matrigel. Cells were staining with the Diffi-Quick Staining kit. B) Heat Maps demonstrating increased (red) or decreased (green) expression of a select number genes from the invasive compared to non-invasive cells. Samples were hybridized to an Agilent whole genome gene expression array following manufacturer’s directions. Arrays were scanned using a GenePix 4000B scanner (Molecular Devices, Sunnyvale, CA) and analyzed using Cluster and Treeview offered by Michael B. Eisen as freeware ( http://rana.lbl.gov/EisenSoftware.htm ). The complete list of genes whose expression changed ≥1.8- or ≤1.8-fold is available in Supplemental Table S1 . C) A select number of up-regulated genes were verified using qRT-PCR for increased expression in the invasive cells. Gray bars represent the non-invasive cell expression normalized to 1 and black bars are relative fold-induction of mRNA from invasive cells. Fold induction was calculated using the Delta-Delta CT method where the non-invasive cells were set at 1.0 as the control, and 18S rRNA was used as a loading control. Data is shown as transformed log2. A Twoway ANOVA with a Bonferroni post-test was performed to compare groups and * represents a p-value of
    Figure Legend Snippet: Gene expression changes in invasive hMSCs A) Microscopic images (20X) of cells invaded toward SCM after 24 hours using either control inserts or inserts containing Matrigel. Cells were staining with the Diffi-Quick Staining kit. B) Heat Maps demonstrating increased (red) or decreased (green) expression of a select number genes from the invasive compared to non-invasive cells. Samples were hybridized to an Agilent whole genome gene expression array following manufacturer’s directions. Arrays were scanned using a GenePix 4000B scanner (Molecular Devices, Sunnyvale, CA) and analyzed using Cluster and Treeview offered by Michael B. Eisen as freeware ( http://rana.lbl.gov/EisenSoftware.htm ). The complete list of genes whose expression changed ≥1.8- or ≤1.8-fold is available in Supplemental Table S1 . C) A select number of up-regulated genes were verified using qRT-PCR for increased expression in the invasive cells. Gray bars represent the non-invasive cell expression normalized to 1 and black bars are relative fold-induction of mRNA from invasive cells. Fold induction was calculated using the Delta-Delta CT method where the non-invasive cells were set at 1.0 as the control, and 18S rRNA was used as a loading control. Data is shown as transformed log2. A Twoway ANOVA with a Bonferroni post-test was performed to compare groups and * represents a p-value of

    Techniques Used: Expressing, Staining, Quantitative RT-PCR, Transformation Assay

    38) Product Images from "MiR-1180-5p regulates apoptosis of Wilms’ tumor by targeting p73"

    Article Title: MiR-1180-5p regulates apoptosis of Wilms’ tumor by targeting p73

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S148684

    MiR-1180 is upregulated in WT, and the inhibition of miR-1180 suppressed WT cell proliferation. Notes: ( A ) The expression of miR-1180 in WT tissues compared with their adjacent non-cancerous tissues (n=30). ( B ) Transfection efficiency measured by the luciferase assay in the indicated cells. ( C ) qRT-PCR analysis of miR-1180 expression in the indicated cells. ( D ) The effect of miR-1180 inhibitor on the viability of WT as measured by the MTT assay. ( E ) Representative crystal violet-stained cell colonies formed by SK-NEP-1 cell lines, 8 days after inoculation. ( F ) The quantification of colonies of the SK-NEP-1 cells tested with colony formation. Data are presented as mean ± SD from three independent experiments with triple replicates per experiment. * p
    Figure Legend Snippet: MiR-1180 is upregulated in WT, and the inhibition of miR-1180 suppressed WT cell proliferation. Notes: ( A ) The expression of miR-1180 in WT tissues compared with their adjacent non-cancerous tissues (n=30). ( B ) Transfection efficiency measured by the luciferase assay in the indicated cells. ( C ) qRT-PCR analysis of miR-1180 expression in the indicated cells. ( D ) The effect of miR-1180 inhibitor on the viability of WT as measured by the MTT assay. ( E ) Representative crystal violet-stained cell colonies formed by SK-NEP-1 cell lines, 8 days after inoculation. ( F ) The quantification of colonies of the SK-NEP-1 cells tested with colony formation. Data are presented as mean ± SD from three independent experiments with triple replicates per experiment. * p

    Techniques Used: Inhibition, Expressing, Transfection, Luciferase, Quantitative RT-PCR, MTT Assay, Staining

    39) Product Images from "LncRNA PVT1 triggers Cyto-protective autophagy and promotes pancreatic ductal adenocarcinoma development via the miR-20a-5p/ULK1 Axis"

    Article Title: LncRNA PVT1 triggers Cyto-protective autophagy and promotes pancreatic ductal adenocarcinoma development via the miR-20a-5p/ULK1 Axis

    Journal: Molecular Cancer

    doi: 10.1186/s12943-018-0845-6

    PVT1 modulates ULK1 expression by sponging miR-20a-5p. a The venn diagram of common microRNAs targeting PVT1 3’UTR and ULK1 3’UTR. Three microRNAs were identified, including miR-20a-5p, miR-302a-3p and miR-17-5p; b PVT1 biotin pull down assay was performed. The level of target microRNAs in the pull down of biotin-labelled PVT1 or negative control was investigated and quantified by qRT-PCR in HPAF-II cells; c Western blot analysis of ULK1 expression in HPAF-II cells with or without overexpression (upper) or suppression (lower) of target microRNAs (miR-20a-5p, miR-302a-3p, miR-17-5p). Interestingly, overexpression of miR-20a-5p suppressed ULK1 protein expression most. While inhibition of miR-20a-5p promoted ULK1 protein expression effectively; d RIP assay and qRT-PCR were conducted to detect the enrichment of PVT1 and miR-20a-5p by using AGO2 antibody in HPAF-II cells. Non-immune IgG acted as an internal control. U6 was used as a non-specific control. Enhanced enrichment of PVT1 and miR-20a-5p with AGO2 antibody was observed compared with IgG; e Bioinformatics prediction of miR-20a-5p binding sites in PVT1 3’UTR sequence using Starbase or ULK1 3’UTR sequence using TargetScan was presented; f Dual-luciferase assays showed diminished luciferase activity was observed when co-transfection of psiCHECK-ULK1-WT and miR-20a-5p occurred in HEK 293 T and HPAF-II cells; g Luciferase reporter assay implied that decreased luciferase activity occurred in HEK 293 T and HPAF-II cells co-transfected with psiCHECK-PVT1-WT and miR-20a-5p compared with luciferase reporter with mutant type of PVT1 and miR-20a-5p; h , i qRT-PCR analysis of miR-20a-5p with augment ( h ) of PVT1 expression in Capan-2 and MIA PaCa-2 cells or attenuation ( i ) of PVT1 expression in SW1990 and HPAF-II cells; j Overexpression of miR-20a-5p made no contribution to the level of PVT1 expression. Data are presented as the mean ± S.D. ( n = 3). * P
    Figure Legend Snippet: PVT1 modulates ULK1 expression by sponging miR-20a-5p. a The venn diagram of common microRNAs targeting PVT1 3’UTR and ULK1 3’UTR. Three microRNAs were identified, including miR-20a-5p, miR-302a-3p and miR-17-5p; b PVT1 biotin pull down assay was performed. The level of target microRNAs in the pull down of biotin-labelled PVT1 or negative control was investigated and quantified by qRT-PCR in HPAF-II cells; c Western blot analysis of ULK1 expression in HPAF-II cells with or without overexpression (upper) or suppression (lower) of target microRNAs (miR-20a-5p, miR-302a-3p, miR-17-5p). Interestingly, overexpression of miR-20a-5p suppressed ULK1 protein expression most. While inhibition of miR-20a-5p promoted ULK1 protein expression effectively; d RIP assay and qRT-PCR were conducted to detect the enrichment of PVT1 and miR-20a-5p by using AGO2 antibody in HPAF-II cells. Non-immune IgG acted as an internal control. U6 was used as a non-specific control. Enhanced enrichment of PVT1 and miR-20a-5p with AGO2 antibody was observed compared with IgG; e Bioinformatics prediction of miR-20a-5p binding sites in PVT1 3’UTR sequence using Starbase or ULK1 3’UTR sequence using TargetScan was presented; f Dual-luciferase assays showed diminished luciferase activity was observed when co-transfection of psiCHECK-ULK1-WT and miR-20a-5p occurred in HEK 293 T and HPAF-II cells; g Luciferase reporter assay implied that decreased luciferase activity occurred in HEK 293 T and HPAF-II cells co-transfected with psiCHECK-PVT1-WT and miR-20a-5p compared with luciferase reporter with mutant type of PVT1 and miR-20a-5p; h , i qRT-PCR analysis of miR-20a-5p with augment ( h ) of PVT1 expression in Capan-2 and MIA PaCa-2 cells or attenuation ( i ) of PVT1 expression in SW1990 and HPAF-II cells; j Overexpression of miR-20a-5p made no contribution to the level of PVT1 expression. Data are presented as the mean ± S.D. ( n = 3). * P

    Techniques Used: Expressing, Pull Down Assay, Negative Control, Quantitative RT-PCR, Western Blot, Over Expression, Inhibition, Binding Assay, Sequencing, Luciferase, Activity Assay, Cotransfection, Reporter Assay, Transfection, Mutagenesis

    ULK1 protein level parallels to that of PVT1 in a subset of human PDA tissues. a Western blot analysis of the expression of ULK1 protein in PDA tissues ( n = 20) and the corresponding adjacent non-tumor pancreatic specimens (n = 20). GAPDH acted as an endogenous control. T, tumor tissues; N, non-tumor pancreatic tissues; b The quantitation of the western blot results in (a). ULK1 protein expression was up-regulated in 13 PDA tissues compared with the corresponding adjacent non-tumor pancreatic specimens in 20 PDA tissues. The high value of ULK1 protein was defined as fold change > 1 ( n = 13); c The level of ULK1 mRNA in 20 PDA tissues was of no statistical significance compared to that of the corresponding adjacent non-tumor pancreatic specimens by qRT-PCR. Data are presented as the mean ± S.D. (n = 20); d PVT1 expression in PDA tissues from the 20 PDA cases based on qRT-PCR analysis. The high value of PVT1 was defined as fold change > 2 ( n = 15), the rest including down-regulation or no evident difference in expression in PDA tissues compared with PDA expression in the corresponding adjacent non-tumor tissue, was defined as low values ( n = 5); e PVT1 expression in PDA tissues based on ISH analysis and the percent of PVT1-positive samples in different groups; f The expression level of PVT1 mRNA was positively correlated with that of ULK1 protein in 20 PDA tissues. The statistical analysis was performed using Pearson’s correlation coefficient ( R = 0.8659, P
    Figure Legend Snippet: ULK1 protein level parallels to that of PVT1 in a subset of human PDA tissues. a Western blot analysis of the expression of ULK1 protein in PDA tissues ( n = 20) and the corresponding adjacent non-tumor pancreatic specimens (n = 20). GAPDH acted as an endogenous control. T, tumor tissues; N, non-tumor pancreatic tissues; b The quantitation of the western blot results in (a). ULK1 protein expression was up-regulated in 13 PDA tissues compared with the corresponding adjacent non-tumor pancreatic specimens in 20 PDA tissues. The high value of ULK1 protein was defined as fold change > 1 ( n = 13); c The level of ULK1 mRNA in 20 PDA tissues was of no statistical significance compared to that of the corresponding adjacent non-tumor pancreatic specimens by qRT-PCR. Data are presented as the mean ± S.D. (n = 20); d PVT1 expression in PDA tissues from the 20 PDA cases based on qRT-PCR analysis. The high value of PVT1 was defined as fold change > 2 ( n = 15), the rest including down-regulation or no evident difference in expression in PDA tissues compared with PDA expression in the corresponding adjacent non-tumor tissue, was defined as low values ( n = 5); e PVT1 expression in PDA tissues based on ISH analysis and the percent of PVT1-positive samples in different groups; f The expression level of PVT1 mRNA was positively correlated with that of ULK1 protein in 20 PDA tissues. The statistical analysis was performed using Pearson’s correlation coefficient ( R = 0.8659, P

    Techniques Used: Western Blot, Expressing, Quantitation Assay, Quantitative RT-PCR, In Situ Hybridization

    40) Product Images from "The clinicopathological significance of HES1 promoter hypomethylation in patients with colorectal cancer"

    Article Title: The clinicopathological significance of HES1 promoter hypomethylation in patients with colorectal cancer

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S151857

    HES1 expression in CRC tissues and adjacent normal samples. Notes: ( A ) The expression of HES1 in 50 cases of CRC tissues and adjacent normal samples was determined using qRT-PCR. Data are presented as mean ± SD of three independent experiments. GAPDH was used as an internal control. * P
    Figure Legend Snippet: HES1 expression in CRC tissues and adjacent normal samples. Notes: ( A ) The expression of HES1 in 50 cases of CRC tissues and adjacent normal samples was determined using qRT-PCR. Data are presented as mean ± SD of three independent experiments. GAPDH was used as an internal control. * P

    Techniques Used: Expressing, Quantitative RT-PCR

    41) Product Images from "Hepatitis B virus X protein promotes human hepatoma cell growth via upregulation of transcription factor AP2α and sphingosine kinase 1"

    Article Title: Hepatitis B virus X protein promotes human hepatoma cell growth via upregulation of transcription factor AP2α and sphingosine kinase 1

    Journal: Acta Pharmacologica Sinica

    doi: 10.1038/aps.2015.38

    The mRNA levels of HBx are positively associated with those of SPHK1 in clinical HCC tissues. (A) The correlation between HBx mRNA levels and SPHK1 mRNA levels was detected by qRT-PCR in clinical HCC tissues ( P
    Figure Legend Snippet: The mRNA levels of HBx are positively associated with those of SPHK1 in clinical HCC tissues. (A) The correlation between HBx mRNA levels and SPHK1 mRNA levels was detected by qRT-PCR in clinical HCC tissues ( P

    Techniques Used: Quantitative RT-PCR

    42) Product Images from "Dynamic Regulation of miRNA Expression by Functionally Enhanced Placental Mesenchymal Stem Cells PromotesHepatic Regeneration in a Rat Model with Bile Duct Ligation"

    Article Title: Dynamic Regulation of miRNA Expression by Functionally Enhanced Placental Mesenchymal Stem Cells PromotesHepatic Regeneration in a Rat Model with Bile Duct Ligation

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20215299

    Improved vascular remodeling by PRL-1(+) PD-MSCs through the regulation of miRNA expression by platelet-derived growth factor receptor alpha (PDGFRA)in a BDL-injured rat model. ( A ) qRT-PCR of endoglin (ENG), and ( B ) platelet-derived growth factor receptor beta (PDGFRB) and ( C ) PDGFR alpha (PDGFRA)-targeted rno-miR-27a-3p expression in BDL-injured rat liver tissue after the administration of naïveand PRL-1(+) PD-MSCs at one, two, three, and fiveweeks. ( D ) Localizationof PDGFRA expression and ( E ) quantification of PDGFRA+ cells in rat liver sections from each group (Con, NTx, TTx Naïve, and TTx PRL-1(+)) at one week as determined by immunofluorescence (red, PDGFRA; blue, DAPI). Scale bars = 100 μm. All experiments wereconducted in at least triplicate. Data from each group are expressed as means ± SD, determined by Student’s t -test.* p
    Figure Legend Snippet: Improved vascular remodeling by PRL-1(+) PD-MSCs through the regulation of miRNA expression by platelet-derived growth factor receptor alpha (PDGFRA)in a BDL-injured rat model. ( A ) qRT-PCR of endoglin (ENG), and ( B ) platelet-derived growth factor receptor beta (PDGFRB) and ( C ) PDGFR alpha (PDGFRA)-targeted rno-miR-27a-3p expression in BDL-injured rat liver tissue after the administration of naïveand PRL-1(+) PD-MSCs at one, two, three, and fiveweeks. ( D ) Localizationof PDGFRA expression and ( E ) quantification of PDGFRA+ cells in rat liver sections from each group (Con, NTx, TTx Naïve, and TTx PRL-1(+)) at one week as determined by immunofluorescence (red, PDGFRA; blue, DAPI). Scale bars = 100 μm. All experiments wereconducted in at least triplicate. Data from each group are expressed as means ± SD, determined by Student’s t -test.* p

    Techniques Used: Expressing, Derivative Assay, Quantitative RT-PCR, Immunofluorescence

    miRNA expression regulates integrin family forPRL-1(+) PD-MSC homingin vivo in a rat model of BDL. ( A ) PKH67 (green)-labeled transplanted cells inBDL-injured rat liver by fluorescence microscopy at one week (arrow = PKH67+ green signals) (blue = 4’,6-diamidino-2-phenylindole; DAPI). mRNA expression levels of ( B ) human-specificAlu sequenceafter the engraftment of naïve (TTxNaïve) and PRL-1(+) PD-MSCs (TTx PRL-1(+)) into injured rat liver compared withthe sham control (Con) and BDL-injurednon-transplantation groups (NTx) at one, two, three, and fiveweeksas determined by qRT-PCR. mRNA and protein levels of ( C ) ITGA4 and ITGB7 and ( D ) integrin alpha 6 (ITGA6)-targeted rno-miR-30a-5p and ( E ) integrin beta 1 (ITGB1)-targeted rno-miR-340-5p expression in rat liver with BDL at one, two, three, and fiveweekspost-transplantation as determined by qRT-PCR.All experiments wereperformed in at least triplicate. Data from each group are shown as means ± SD, determinedby Student’s t -test. Scale bars = 100 μm.* p
    Figure Legend Snippet: miRNA expression regulates integrin family forPRL-1(+) PD-MSC homingin vivo in a rat model of BDL. ( A ) PKH67 (green)-labeled transplanted cells inBDL-injured rat liver by fluorescence microscopy at one week (arrow = PKH67+ green signals) (blue = 4’,6-diamidino-2-phenylindole; DAPI). mRNA expression levels of ( B ) human-specificAlu sequenceafter the engraftment of naïve (TTxNaïve) and PRL-1(+) PD-MSCs (TTx PRL-1(+)) into injured rat liver compared withthe sham control (Con) and BDL-injurednon-transplantation groups (NTx) at one, two, three, and fiveweeksas determined by qRT-PCR. mRNA and protein levels of ( C ) ITGA4 and ITGB7 and ( D ) integrin alpha 6 (ITGA6)-targeted rno-miR-30a-5p and ( E ) integrin beta 1 (ITGB1)-targeted rno-miR-340-5p expression in rat liver with BDL at one, two, three, and fiveweekspost-transplantation as determined by qRT-PCR.All experiments wereperformed in at least triplicate. Data from each group are shown as means ± SD, determinedby Student’s t -test. Scale bars = 100 μm.* p

    Techniques Used: Expressing, Labeling, Fluorescence, Microscopy, Transplantation Assay, Quantitative RT-PCR

    PRL-1-Targeted miRNA expression regulates migration ability through the RHO family. ( A ) Representative images and ( B ) the number of migrated cells in the naïve PD-MSCs and PRL-1(+) PD-MSCs determined using a Transwell insert system following small interfering RNA (siRNA) PRL-1 (siPRL-1) treatment (50 nM) for 24 h. ( C ) mRNA expression of PRL-1 and targeted hsa-miR-30a-5p expression, and ( D ) RHOA and ROCK1 in migratednaïve and PRL-1(+) PD-MSCs determined using a Transwell insert system following siPRL-1 treatment (50 nM) for 24 h as determined by qRT-PCR.( E ) Protein levels of RHOA and ROCK1 and ( F ) their quantification in migrated naïve and PRL-1(+) PD-MSCs according to siPRL-1 treatment by Western blotting. All experiments were performed in at least triplicate. Data from each group are shown as means ± SD, determined by Student’s t -test. Scale bars = 100 μm. * p
    Figure Legend Snippet: PRL-1-Targeted miRNA expression regulates migration ability through the RHO family. ( A ) Representative images and ( B ) the number of migrated cells in the naïve PD-MSCs and PRL-1(+) PD-MSCs determined using a Transwell insert system following small interfering RNA (siRNA) PRL-1 (siPRL-1) treatment (50 nM) for 24 h. ( C ) mRNA expression of PRL-1 and targeted hsa-miR-30a-5p expression, and ( D ) RHOA and ROCK1 in migratednaïve and PRL-1(+) PD-MSCs determined using a Transwell insert system following siPRL-1 treatment (50 nM) for 24 h as determined by qRT-PCR.( E ) Protein levels of RHOA and ROCK1 and ( F ) their quantification in migrated naïve and PRL-1(+) PD-MSCs according to siPRL-1 treatment by Western blotting. All experiments were performed in at least triplicate. Data from each group are shown as means ± SD, determined by Student’s t -test. Scale bars = 100 μm. * p

    Techniques Used: Expressing, Migration, Small Interfering RNA, Quantitative RT-PCR, Western Blot

    Phosphatase of regenerating liver-1 (PRL-1)-dependent migration ability under hypoxic conditions regulated by miRNAs targeting the integrin family.( A ) Representative images and ( B ) the number of migrated naïve and PRL-1(+) PD-MSCs using a Transwell insert system under 1% hypoxic conditions for 24 h. ( C ) mRNA expression of hypoxia-inducible factor 1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF) in migrated PRL-1(+) PD-MSCs determined using a Transwell insert system under 1% hypoxic conditions for 24 h, as determinedby qRT-PCR. ( D ) Ras homolog family member A (RHOA) and RHO-associated coiled-coil-containing protein kinase 1 (ROCK1) levelsin migratednaïve and PRL-1(+) PD-MSCs under 1% hypoxic conditions as determined by qRT-PCR. ( E ) mRNA expression levels of PRL-1 and targeted hsa-miR-30a-5p expression, ( F ) Integrin alpha 4 (ITGA4) and targeted hsa-miR-340-5p expression, and ( G ) integrin beta 7 (ITGB7) and targeted hsa-miR-146a-3p expression in migrated PRL-1(+) PD-MSCs under 1% hypoxic conditions for 24 h as determined by qRT-PCR. Luciferase assay of PRL-1, ITGA4, and ITGB7 containing hsa-miR-30a-5p, 340-5p, and 146a-3p binding sitesin ( H ) naïve and ( I ) PRL-1(+) PD-MSCs. Firefly luciferase activities were measured by luminescence. All experiments were conducted in at least triplicate. Data from each group are expressed as means ± SD, determined by Student’s t -test. Scale bars = 100 μm.* p
    Figure Legend Snippet: Phosphatase of regenerating liver-1 (PRL-1)-dependent migration ability under hypoxic conditions regulated by miRNAs targeting the integrin family.( A ) Representative images and ( B ) the number of migrated naïve and PRL-1(+) PD-MSCs using a Transwell insert system under 1% hypoxic conditions for 24 h. ( C ) mRNA expression of hypoxia-inducible factor 1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF) in migrated PRL-1(+) PD-MSCs determined using a Transwell insert system under 1% hypoxic conditions for 24 h, as determinedby qRT-PCR. ( D ) Ras homolog family member A (RHOA) and RHO-associated coiled-coil-containing protein kinase 1 (ROCK1) levelsin migratednaïve and PRL-1(+) PD-MSCs under 1% hypoxic conditions as determined by qRT-PCR. ( E ) mRNA expression levels of PRL-1 and targeted hsa-miR-30a-5p expression, ( F ) Integrin alpha 4 (ITGA4) and targeted hsa-miR-340-5p expression, and ( G ) integrin beta 7 (ITGB7) and targeted hsa-miR-146a-3p expression in migrated PRL-1(+) PD-MSCs under 1% hypoxic conditions for 24 h as determined by qRT-PCR. Luciferase assay of PRL-1, ITGA4, and ITGB7 containing hsa-miR-30a-5p, 340-5p, and 146a-3p binding sitesin ( H ) naïve and ( I ) PRL-1(+) PD-MSCs. Firefly luciferase activities were measured by luminescence. All experiments were conducted in at least triplicate. Data from each group are expressed as means ± SD, determined by Student’s t -test. Scale bars = 100 μm.* p

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

    MicroRNA (miRNA) profiling of migrated naïve placenta-derived mesenchymal stem cells (PD-MSCs) under hypoxic conditions and in bile duct ligation (BDL)-injured liver in rats. ( A ) Messenger RNA (mRNA)expression of hypoxia-inducible factor 1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF) in migrated naïve PD-MSCs determined using a Transwell insert system under 1% hypoxic conditions for 24 h, as determinedby quantitative real-time polymerase chain reaction (qRT-PCR). ( B ) Heat map and ( C ) Venn diagram of the microarray results ofmigrated naïve PD-MSCs under hypoxic conditions compared with normoxic conditions (Hyp/Nor), transplanted naïve (TTxNaïve) compared to NTx at one week (TTx/NTx 1w), and TTx Naïve compared to NTx at two weeks (TTx/NTx 2w). qRT-PCR runs were conducted in at least triplicate. Data from each group are shown as means ± SD, determined by Student’s t -test; * p
    Figure Legend Snippet: MicroRNA (miRNA) profiling of migrated naïve placenta-derived mesenchymal stem cells (PD-MSCs) under hypoxic conditions and in bile duct ligation (BDL)-injured liver in rats. ( A ) Messenger RNA (mRNA)expression of hypoxia-inducible factor 1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF) in migrated naïve PD-MSCs determined using a Transwell insert system under 1% hypoxic conditions for 24 h, as determinedby quantitative real-time polymerase chain reaction (qRT-PCR). ( B ) Heat map and ( C ) Venn diagram of the microarray results ofmigrated naïve PD-MSCs under hypoxic conditions compared with normoxic conditions (Hyp/Nor), transplanted naïve (TTxNaïve) compared to NTx at one week (TTx/NTx 1w), and TTx Naïve compared to NTx at two weeks (TTx/NTx 2w). qRT-PCR runs were conducted in at least triplicate. Data from each group are shown as means ± SD, determined by Student’s t -test; * p

    Techniques Used: Derivative Assay, Ligation, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Microarray

    miRNAs mediated hepatic regeneration by PRL-1(+) PD-MSCs in a rat model of BDL through interleukin-6 (IL-6)/signal transducer and activator of transcription 3 (STAT3) signaling. ( A ) qRT-PCR of rno-miR-21-5p-targeted interleukin-6 receptor (IL-6R) and ( B ) Western blotting of glycoprotein 130 (gp130), IL-6, and phosphorylated STAT3 levels in BDL-injured rat liver tissue following the administration of naïve and PRL-1(+) PD-MSCs at one, two, three, and fiveweeks. ( C ) mRNA expression of hepatocyte nuclear factor 4 alpha (HNF4A) and ( D ) miR-122-5p-targeted HNF1 homeobox A (HNF1A) in BDL-injured rat liver tissue by qRT-PCR. ( E ) Proliferating cell nuclear antigen (PCNA) expression in the rat liver sections from each group (Con, NTx, TTxNaïve, and TTxPRL-1(+)) at one week as determined by immunohistochemistry. ( F ) Quantification of the PCNA-positive area in hepatocytes. Scale bars = 50μm. All experiments wereconductedin at least triplicate.Data from each group are expressed as means ± SD, determined by Student’s t -test.* p
    Figure Legend Snippet: miRNAs mediated hepatic regeneration by PRL-1(+) PD-MSCs in a rat model of BDL through interleukin-6 (IL-6)/signal transducer and activator of transcription 3 (STAT3) signaling. ( A ) qRT-PCR of rno-miR-21-5p-targeted interleukin-6 receptor (IL-6R) and ( B ) Western blotting of glycoprotein 130 (gp130), IL-6, and phosphorylated STAT3 levels in BDL-injured rat liver tissue following the administration of naïve and PRL-1(+) PD-MSCs at one, two, three, and fiveweeks. ( C ) mRNA expression of hepatocyte nuclear factor 4 alpha (HNF4A) and ( D ) miR-122-5p-targeted HNF1 homeobox A (HNF1A) in BDL-injured rat liver tissue by qRT-PCR. ( E ) Proliferating cell nuclear antigen (PCNA) expression in the rat liver sections from each group (Con, NTx, TTxNaïve, and TTxPRL-1(+)) at one week as determined by immunohistochemistry. ( F ) Quantification of the PCNA-positive area in hepatocytes. Scale bars = 50μm. All experiments wereconductedin at least triplicate.Data from each group are expressed as means ± SD, determined by Student’s t -test.* p

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

    43) Product Images from "Long non-coding RNA HULC promotes tumor angiogenesis in liver cancer by up-regulating sphingosine kinase 1 (SPHK1)"

    Article Title: Long non-coding RNA HULC promotes tumor angiogenesis in liver cancer by up-regulating sphingosine kinase 1 (SPHK1)

    Journal: Oncotarget

    doi:

    HULC increases E2F1 by sequestering miR-107 A. Relative luciferase activity of pGL3-E2F1 was measured by luciferase reporter gene assays in HepG2 cells transfected with miR-107 or co-transfected with miR-107 inhibitor. B. Relative luciferase activities of pGL3-E2F1-mut were detected in HepG2 and H7402 cells treated with miR-107. C. E2F1 was measured by Western blot analysis in HepG2 and H7402 cells transfected with miR-107 or co-transfected with miR-107 and miR-107 inhibitor. D. A model shows the predicted interaction between HULC and miR-107 through complementary base-pairs. The generated mutant site at the HULC (365-383) is indicated. E. Relative luciferase activities of pGL3-E2F1 were measured by luciferase reporter gene assays in HepG2 cells transfected with miR-107 or co-transfected with HULC (or HULC-107-mut). F. The correlation between HULC mRNA levels and miR-107 levels was detected by qRT-PCR in 60 HCC tissues ( P
    Figure Legend Snippet: HULC increases E2F1 by sequestering miR-107 A. Relative luciferase activity of pGL3-E2F1 was measured by luciferase reporter gene assays in HepG2 cells transfected with miR-107 or co-transfected with miR-107 inhibitor. B. Relative luciferase activities of pGL3-E2F1-mut were detected in HepG2 and H7402 cells treated with miR-107. C. E2F1 was measured by Western blot analysis in HepG2 and H7402 cells transfected with miR-107 or co-transfected with miR-107 and miR-107 inhibitor. D. A model shows the predicted interaction between HULC and miR-107 through complementary base-pairs. The generated mutant site at the HULC (365-383) is indicated. E. Relative luciferase activities of pGL3-E2F1 were measured by luciferase reporter gene assays in HepG2 cells transfected with miR-107 or co-transfected with HULC (or HULC-107-mut). F. The correlation between HULC mRNA levels and miR-107 levels was detected by qRT-PCR in 60 HCC tissues ( P

    Techniques Used: Luciferase, Activity Assay, Transfection, Western Blot, Generated, Mutagenesis, Quantitative RT-PCR

    HULC is able to increase the expression of E2F1 A. , B. The expression of E2F1 was examined by Western blot analysis in HepG2 and H7402 (or HepG2.2.15) cells transfected with pcDNA3.1-HULC (or si-HULC-1). The transfection efficiency of HULC (or si-HULC-1) was detected by qRT-PCR. C. The correlation between E2F1 mRNA levels and HULC mRNA levels was detected by qRT-PCR in 60 HCC tissues ( P
    Figure Legend Snippet: HULC is able to increase the expression of E2F1 A. , B. The expression of E2F1 was examined by Western blot analysis in HepG2 and H7402 (or HepG2.2.15) cells transfected with pcDNA3.1-HULC (or si-HULC-1). The transfection efficiency of HULC (or si-HULC-1) was detected by qRT-PCR. C. The correlation between E2F1 mRNA levels and HULC mRNA levels was detected by qRT-PCR in 60 HCC tissues ( P

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

    HULC contributes to angiogenesis in liver cancer by up-regulating SPHK1 A. The correlation between SPHK1 mRNA levels and HULC mRNA levels was detected by qRT-PCR in 60 HCC tissues ( P
    Figure Legend Snippet: HULC contributes to angiogenesis in liver cancer by up-regulating SPHK1 A. The correlation between SPHK1 mRNA levels and HULC mRNA levels was detected by qRT-PCR in 60 HCC tissues ( P

    Techniques Used: Quantitative RT-PCR

    44) Product Images from "PinX1 suppresses bladder urothelial carcinoma cell proliferation via the inhibition of telomerase activity and p16/cyclin D1 pathway"

    Article Title: PinX1 suppresses bladder urothelial carcinoma cell proliferation via the inhibition of telomerase activity and p16/cyclin D1 pathway

    Journal: Molecular Cancer

    doi: 10.1186/1476-4598-12-148

    The expression of PinX1 in UCB and adjacent normal bladder tissues. (A) Down-regulated expression of PinX1 mRNA was examined by qRT-PCR in 8/10 UCB cases, when compared with adjacent normal bladder tissues. Expression levels were normalized for GAPDH. Error bars, SD calculated from three parallel experiments. (B) Down-regulated expression of PinX1 protein was detected by Western blotting in 7/10 UCB cases, when compared with adjacent normal bladder tissues. Expression levels were normalized with GAPDH. (C-F) The expression of PinX1 in UCB and adjacent normal bladder tissues by IHC (100×). An UCB (case 45) tissue showed negative expression of PinX1 (C) , while its adjacent normal bladder urothelial mucosal tissue was positive stained by PinX1, in which more than 90% of tumor cells were positively stained by PinX1 in the nucleus (D) . Negative expression of PinX1 was observed in another UCB tissue (case 73), in which only 10% of tumor cells demonstrated a nuclear staining of PinX1 (E) . An UCB (case 126) was negatively stained by PinX1 (F) .
    Figure Legend Snippet: The expression of PinX1 in UCB and adjacent normal bladder tissues. (A) Down-regulated expression of PinX1 mRNA was examined by qRT-PCR in 8/10 UCB cases, when compared with adjacent normal bladder tissues. Expression levels were normalized for GAPDH. Error bars, SD calculated from three parallel experiments. (B) Down-regulated expression of PinX1 protein was detected by Western blotting in 7/10 UCB cases, when compared with adjacent normal bladder tissues. Expression levels were normalized with GAPDH. (C-F) The expression of PinX1 in UCB and adjacent normal bladder tissues by IHC (100×). An UCB (case 45) tissue showed negative expression of PinX1 (C) , while its adjacent normal bladder urothelial mucosal tissue was positive stained by PinX1, in which more than 90% of tumor cells were positively stained by PinX1 in the nucleus (D) . Negative expression of PinX1 was observed in another UCB tissue (case 73), in which only 10% of tumor cells demonstrated a nuclear staining of PinX1 (E) . An UCB (case 126) was negatively stained by PinX1 (F) .

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

    45) Product Images from "The Effect of 3D Nanofibrous Scaffolds on the Chondrogenesis of Induced Pluripotent Stem Cells and Their Application in Restoration of Cartilage Defects"

    Article Title: The Effect of 3D Nanofibrous Scaffolds on the Chondrogenesis of Induced Pluripotent Stem Cells and Their Application in Restoration of Cartilage Defects

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0111566

    Chondrogenesis of iPSCs cultured on scaffolds (scaffold group) or plates (control group) in vitro . Collagen II synthesis in the scaffold group ( A,B,C ) and control group ( D,E,F ) was analyzed by immunofluorescence staining, including nuclear counterstaining with DAPI ( A,D ), anti-collagen II staining in red ( B,E ); merged images are also shown ( C,F ). ( G ) qRT-PCR analysis of gene expression and ( H ) western blotting for collagen II, aggrecan, sox9 and collagen I, using GAPDH as the internal reference. The nanofibrous scaffolds generally enhanced cartilage-specific gene expression and protein levels. ♦ Statistically significant difference compared with the control group ( p
    Figure Legend Snippet: Chondrogenesis of iPSCs cultured on scaffolds (scaffold group) or plates (control group) in vitro . Collagen II synthesis in the scaffold group ( A,B,C ) and control group ( D,E,F ) was analyzed by immunofluorescence staining, including nuclear counterstaining with DAPI ( A,D ), anti-collagen II staining in red ( B,E ); merged images are also shown ( C,F ). ( G ) qRT-PCR analysis of gene expression and ( H ) western blotting for collagen II, aggrecan, sox9 and collagen I, using GAPDH as the internal reference. The nanofibrous scaffolds generally enhanced cartilage-specific gene expression and protein levels. ♦ Statistically significant difference compared with the control group ( p

    Techniques Used: Cell Culture, In Vitro, Immunofluorescence, Staining, Quantitative RT-PCR, Expressing, Western Blot

    Gene expression and protein levels in vivo . Collagen I ( A ) and collagen II ( B ) gene expression levels over 3 months were determined using qRT-PCR, and western blotting was used to evaluate the collagen II, aggrecan, sox9 and collagen I protein levels in the regeneration area ( C ). The scaffold notably stimulated the expression of the cartilage-related markers, but not collagen I. ♦ Statistically significant difference compared with the control group ( p
    Figure Legend Snippet: Gene expression and protein levels in vivo . Collagen I ( A ) and collagen II ( B ) gene expression levels over 3 months were determined using qRT-PCR, and western blotting was used to evaluate the collagen II, aggrecan, sox9 and collagen I protein levels in the regeneration area ( C ). The scaffold notably stimulated the expression of the cartilage-related markers, but not collagen I. ♦ Statistically significant difference compared with the control group ( p

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

    46) Product Images from "TNF-α Induced the Enhanced Apoptosis of Mesenchymal Stem Cells in Ankylosing Spondylitis by Overexpressing TRAIL-R2"

    Article Title: TNF-α Induced the Enhanced Apoptosis of Mesenchymal Stem Cells in Ankylosing Spondylitis by Overexpressing TRAIL-R2

    Journal: Stem Cells International

    doi: 10.1155/2017/4521324

    The DR and mitochondrial pathways were both involved in MSC apoptosis. (a) qRT-PCR was performed to compare the gene expressions of total caspase-8, total cytochrome C, Fas, and TNFR1 between HDMSCs ( n = 28) and ASMSCs ( n = 22). ASMSCs expressed much higher gene expression levels of total caspase-8 than HDMSCs after treatment with TNF- α /CHX, while no difference was found between HDMSCs and ASMSCs in total cytochrome C, Fas, or TNFR1 gene expression levels. (b) A western blot was performed to compare the protein expressions of cleaved caspase-8, cytosolic cytochrome C, TNFR1, and Fas between HDMSCs ( n = 28) and ASMSCs ( n = 22). After treatment with TNF- α /CHX, ASMSCs had higher cleaved caspase-8 and cytosolic cytochrome C protein expression levels than HDMSCs and similar protein levels of TNFR1 and Fas as HDMSCs. Fold changes are presented as the mean ± SD. ∗ indicates P
    Figure Legend Snippet: The DR and mitochondrial pathways were both involved in MSC apoptosis. (a) qRT-PCR was performed to compare the gene expressions of total caspase-8, total cytochrome C, Fas, and TNFR1 between HDMSCs ( n = 28) and ASMSCs ( n = 22). ASMSCs expressed much higher gene expression levels of total caspase-8 than HDMSCs after treatment with TNF- α /CHX, while no difference was found between HDMSCs and ASMSCs in total cytochrome C, Fas, or TNFR1 gene expression levels. (b) A western blot was performed to compare the protein expressions of cleaved caspase-8, cytosolic cytochrome C, TNFR1, and Fas between HDMSCs ( n = 28) and ASMSCs ( n = 22). After treatment with TNF- α /CHX, ASMSCs had higher cleaved caspase-8 and cytosolic cytochrome C protein expression levels than HDMSCs and similar protein levels of TNFR1 and Fas as HDMSCs. Fold changes are presented as the mean ± SD. ∗ indicates P

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

    ASMSCs had increased protein expression of cleaved caspase-3, TRAIL-R2, and FADD and increased gene expression of TRAIL-R2 and FADD during apoptosis. (a) The Proteome Profiler™ Human Apoptosis Array Kit was used to determine the expression levels of 35 apoptosis-related proteins in HDMSCs ( n = 6) and ASMSCs ( n = 6) after treatment with TNF- α /CHX. The intensity of each pair of spots represents repeated measures of one protein. ASMSCs had lower pro-caspase-3 (a.1) expression but higher cleaved caspase-3 (a.2), TRAIL-R2 (a.3), and FADD (a.4) expression levels than HDMSCs. No significant differences in the expression of the remaining 31 proteins were found between HDMSCs and ASMSCs (b) qRT-PCR was performed to confirm the results of the Proteome Profiler™ Human Apoptosis Array. The results of TRAIL-R2 and FADD in HDMSCs ( n = 28) and ASMSCs ( n = 22) were consistent with those of the Proteome Profiler™ Human Apoptosis Array. However, total caspase-3 mRNA expression was not different between HDMSCs and ASMSCs. (c) Western blot analysis was performed on proteins isolated from HDMSCs ( n = 28) and ASMSCs ( n = 22) to further confirm the results of the Proteome Profiler™ Human Apoptosis Array, and the results of both methods were in agreement. Values are presented as the mean ± SD. ∗ indicates P
    Figure Legend Snippet: ASMSCs had increased protein expression of cleaved caspase-3, TRAIL-R2, and FADD and increased gene expression of TRAIL-R2 and FADD during apoptosis. (a) The Proteome Profiler™ Human Apoptosis Array Kit was used to determine the expression levels of 35 apoptosis-related proteins in HDMSCs ( n = 6) and ASMSCs ( n = 6) after treatment with TNF- α /CHX. The intensity of each pair of spots represents repeated measures of one protein. ASMSCs had lower pro-caspase-3 (a.1) expression but higher cleaved caspase-3 (a.2), TRAIL-R2 (a.3), and FADD (a.4) expression levels than HDMSCs. No significant differences in the expression of the remaining 31 proteins were found between HDMSCs and ASMSCs (b) qRT-PCR was performed to confirm the results of the Proteome Profiler™ Human Apoptosis Array. The results of TRAIL-R2 and FADD in HDMSCs ( n = 28) and ASMSCs ( n = 22) were consistent with those of the Proteome Profiler™ Human Apoptosis Array. However, total caspase-3 mRNA expression was not different between HDMSCs and ASMSCs. (c) Western blot analysis was performed on proteins isolated from HDMSCs ( n = 28) and ASMSCs ( n = 22) to further confirm the results of the Proteome Profiler™ Human Apoptosis Array, and the results of both methods were in agreement. Values are presented as the mean ± SD. ∗ indicates P

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

    47) Product Images from "Knockdown of miR-27a sensitizes colorectal cancer stem cells to TRAIL by promoting the formation of Apaf-1-caspase-9 complex"

    Article Title: Knockdown of miR-27a sensitizes colorectal cancer stem cells to TRAIL by promoting the formation of Apaf-1-caspase-9 complex

    Journal: Oncotarget

    doi: 10.18632/oncotarget.16779

    Overexpresion of miR-27a in colorectal cancer stem cells ( A ) Flow cytometry analysis was performed to detect the populations of CSCs and non-CSCs in HT29 and SW48 cell lines. ( B ) Expression of miR-27a in FHC, HT29 and SW480 CSCs and non-CSCs was measured by using qRT-PCR analysis. * P
    Figure Legend Snippet: Overexpresion of miR-27a in colorectal cancer stem cells ( A ) Flow cytometry analysis was performed to detect the populations of CSCs and non-CSCs in HT29 and SW48 cell lines. ( B ) Expression of miR-27a in FHC, HT29 and SW480 CSCs and non-CSCs was measured by using qRT-PCR analysis. * P

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

    48) Product Images from "Identification of aberrantly expressed long non-coding RNAs in stomach adenocarcinoma"

    Article Title: Identification of aberrantly expressed long non-coding RNAs in stomach adenocarcinoma

    Journal: Oncotarget

    doi: 10.18632/oncotarget.17329

    qRT-PCR validation of dysregulated DElncRNAs and DEmRNAs in STAD compared with matched non-tumor tissues ( A ) The expression level of lncRNA LINC00982; ( B ) The expression level of lncRNA SNHG3; ( C ) The expression level of lncRNA LINC00261; ( D ) The expression level of FOXA2. STAD indicated stomach adenocarcinoma; CON indicated paired adjacent non-tumor tissues. * represented P
    Figure Legend Snippet: qRT-PCR validation of dysregulated DElncRNAs and DEmRNAs in STAD compared with matched non-tumor tissues ( A ) The expression level of lncRNA LINC00982; ( B ) The expression level of lncRNA SNHG3; ( C ) The expression level of lncRNA LINC00261; ( D ) The expression level of FOXA2. STAD indicated stomach adenocarcinoma; CON indicated paired adjacent non-tumor tissues. * represented P

    Techniques Used: Quantitative RT-PCR, Expressing

    49) Product Images from "PDZ binding kinase, regulated by FoxM1, enhances malignant phenotype via activation of β-Catenin signaling in hepatocellular carcinoma"

    Article Title: PDZ binding kinase, regulated by FoxM1, enhances malignant phenotype via activation of β-Catenin signaling in hepatocellular carcinoma

    Journal: Oncotarget

    doi: 10.18632/oncotarget.17587

    PBK expression is up-regulated in HCC and associated with poor outcomes ( A ) The expression of PBK mRNA in 56 pairs of HCC specimens was determined by qRT-PCR. ( B ) PBK protein level was evaluated in 16 HCC cases by western blot. The representative images (left panel) and the statistics (right panel) were presented. ( C ) PBK expression was examined 520 paired HCC and nontumorous tissues and 96 paired HCC and metastatic tissues by immunohistochemistry (IHC). The IHC score of PBK was shown and compared. ( D , E ) Kaplan-Meier analyses was conducted to assess the value of PBK in overall and disease-free survivals of patients with HCC in our and TCGA cohorts.
    Figure Legend Snippet: PBK expression is up-regulated in HCC and associated with poor outcomes ( A ) The expression of PBK mRNA in 56 pairs of HCC specimens was determined by qRT-PCR. ( B ) PBK protein level was evaluated in 16 HCC cases by western blot. The representative images (left panel) and the statistics (right panel) were presented. ( C ) PBK expression was examined 520 paired HCC and nontumorous tissues and 96 paired HCC and metastatic tissues by immunohistochemistry (IHC). The IHC score of PBK was shown and compared. ( D , E ) Kaplan-Meier analyses was conducted to assess the value of PBK in overall and disease-free survivals of patients with HCC in our and TCGA cohorts.

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

    50) Product Images from "Could −79 °C Spray-Type Cryotherapy Be an Effective Monotherapy for the Treatment of Keloid?"

    Article Title: Could −79 °C Spray-Type Cryotherapy Be an Effective Monotherapy for the Treatment of Keloid?

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms18122536

    Cryotherapy increases the migration and proliferative activity of keloid dermal fibroblasts. ( a ) The comparison of the proliferative activity of the dermal fibroblasts from keloids before and after cryotherapy. Cell proliferation was promoted in the fibroblast treated with cryotherapy. The cell number was estimated at 48 h after cell seeding; ( b , c ) The migration activity of dermal fibroblasts was increased following cryotherapy. The migration of the cells was analysed by the transwell assay ( b ) and wound-healing assay ( c ); ( d ) qRT-PCR experiments were performed to evaluate the mRNA levels of fibronectin, MMP2 (Matrix metalloproteinase 2), MMP9, PAI-1 (Plasminogen activator inhibitor-1), TGF-β1, and Col2a1 (Collagen Type II Alpha 1) in fibroblasts derived from the dermis under the cryotherapy-treated (+) or untreated (−) keloids. MMP9 and TGF-β1 were significantly increased in the fibroblast treated with cryotherapy as compared with controls; ( e ) Immunoblot analysis of phosphorylated Smad2/3. Total Smad2/3 and GAPDH were used as the loading controls. Error bars represent the standard error from three repeated experiments. * p
    Figure Legend Snippet: Cryotherapy increases the migration and proliferative activity of keloid dermal fibroblasts. ( a ) The comparison of the proliferative activity of the dermal fibroblasts from keloids before and after cryotherapy. Cell proliferation was promoted in the fibroblast treated with cryotherapy. The cell number was estimated at 48 h after cell seeding; ( b , c ) The migration activity of dermal fibroblasts was increased following cryotherapy. The migration of the cells was analysed by the transwell assay ( b ) and wound-healing assay ( c ); ( d ) qRT-PCR experiments were performed to evaluate the mRNA levels of fibronectin, MMP2 (Matrix metalloproteinase 2), MMP9, PAI-1 (Plasminogen activator inhibitor-1), TGF-β1, and Col2a1 (Collagen Type II Alpha 1) in fibroblasts derived from the dermis under the cryotherapy-treated (+) or untreated (−) keloids. MMP9 and TGF-β1 were significantly increased in the fibroblast treated with cryotherapy as compared with controls; ( e ) Immunoblot analysis of phosphorylated Smad2/3. Total Smad2/3 and GAPDH were used as the loading controls. Error bars represent the standard error from three repeated experiments. * p

    Techniques Used: Migration, Activity Assay, Transwell Assay, Wound Healing Assay, Quantitative RT-PCR, Derivative Assay

    51) Product Images from "Downregulation of FOXO6 in breast cancer promotes epithelial–mesenchymal transition and facilitates migration and proliferation of cancer cells"

    Article Title: Downregulation of FOXO6 in breast cancer promotes epithelial–mesenchymal transition and facilitates migration and proliferation of cancer cells

    Journal: Cancer Management and Research

    doi: 10.2147/CMAR.S157661

    FOXO6 is downregulated in breast cancer tissues and associated with EMT-associated proteins E-cadherin and N-cadherin. Notes: ( A ) The expression of FOXO6 in tumor tissues and adjacent normal tissues was detected by qRT-PCR and Western blotting analysis, respectively. Tumor tissues vs adjacent normal tissues, * p
    Figure Legend Snippet: FOXO6 is downregulated in breast cancer tissues and associated with EMT-associated proteins E-cadherin and N-cadherin. Notes: ( A ) The expression of FOXO6 in tumor tissues and adjacent normal tissues was detected by qRT-PCR and Western blotting analysis, respectively. Tumor tissues vs adjacent normal tissues, * p

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

    FOXO6 inhibits breast cancer cells migration and invasion. Notes: ( A ) MDA-MB-231 cells were transfected with vector or FOXO6, or siControl or siFOXO6, respectively. After transfection for 48 h, the expression of FOXO6 was determined by qRT-PCR and Western blotting analyses. FOXO6 vs vector, siFOXO6 vs siControl, * p
    Figure Legend Snippet: FOXO6 inhibits breast cancer cells migration and invasion. Notes: ( A ) MDA-MB-231 cells were transfected with vector or FOXO6, or siControl or siFOXO6, respectively. After transfection for 48 h, the expression of FOXO6 was determined by qRT-PCR and Western blotting analyses. FOXO6 vs vector, siFOXO6 vs siControl, * p

    Techniques Used: Migration, Multiple Displacement Amplification, Transfection, Plasmid Preparation, Expressing, Quantitative RT-PCR, Western Blot

    Sirt6 is a direct transcriptional target of FOXO6. Notes: ( A ) FOXO6 was overexpressed or knocked down in MCF-7 and MDA-MB-231 cells. The mRNA level of Sirt6 was assessed by qRT-PCR assay. FOXO6 vs vector, siFOXO6 vs siControl, * p
    Figure Legend Snippet: Sirt6 is a direct transcriptional target of FOXO6. Notes: ( A ) FOXO6 was overexpressed or knocked down in MCF-7 and MDA-MB-231 cells. The mRNA level of Sirt6 was assessed by qRT-PCR assay. FOXO6 vs vector, siFOXO6 vs siControl, * p

    Techniques Used: Multiple Displacement Amplification, Quantitative RT-PCR, Plasmid Preparation

    FOXO6 suppresses EMT in breast cancer cells. Notes: ( A ) MCF-7 cells were transfected with vector or FOXO6, or siControl or siFOXO6, respectively. After transfection for 48 h, the expression of FOXO6 was determined by qRT-PCR and Western blotting analyses. FOXO6 vs vector, siFOXO6 vs siControl, * p
    Figure Legend Snippet: FOXO6 suppresses EMT in breast cancer cells. Notes: ( A ) MCF-7 cells were transfected with vector or FOXO6, or siControl or siFOXO6, respectively. After transfection for 48 h, the expression of FOXO6 was determined by qRT-PCR and Western blotting analyses. FOXO6 vs vector, siFOXO6 vs siControl, * p

    Techniques Used: Transfection, Plasmid Preparation, Expressing, Quantitative RT-PCR, Western Blot

    52) Product Images from "Elongator promotes the migration and invasion of hepatocellular carcinoma cell by the phosphorylation of AKT"

    Article Title: Elongator promotes the migration and invasion of hepatocellular carcinoma cell by the phosphorylation of AKT

    Journal: International Journal of Biological Sciences

    doi: 10.7150/ijbs.23511

    Elongator activates PI3K/AKT/MMPs signaling pathway. Stably transfected cells were maintained in the absence of serum for 24 h, and then, total RNA or proteins were isolated. Overexpressed Elp3 and Elp4 in HepG2 cells were abbreviated to Elp3o and Elp4o. Elp3i and Elp3i-b were two different clones targeting the same sequence of Elp3 transcript. Elp4i and Elp4i-b were also two different clones targeting the same sequence of Elp4 transcript. ( A ) The mRNA expression of MMP-2 and MMP-9 in stably transfected HepG2 cell lines was examined by qRT-PCR. Related mRNA levels of MMP-2 and MMP-9 were compared between control HepG2 cells and Elp3o or Elp4o cells, and between control HepG2 cells and Elp3i or Elp4i cells. MMP-2 and MMP-9 mRNA levels in control HepG2 cells were set to 1 fold. ( B ) The plasmids for Elp3 overexpression (Elp3o), Elp3 interference (Elp3i), or vector alone (Vector) were transfected into Hep3B cells. The protein expression of Elp3 and Elp4 in transiently transfected Hep3B cells were examined by western blot analysis. GAPDH was shown as a loading control. ( C ) Rescue Experiment. The plasmids for Elp3 overexpression (Elp3o) were transiently transfected into HepG2-Elp3i cells. Cells were subjected to western blot analysis with antibodies against AKT, phosphorylated AKT, MMP-2 and MMP-9. GAPDH was shown as a loading control. ( D-G ) HepG2 cells were transfected with increasing amount of CMV4-Elp3o (D), CMV4-Elp4o (E), GV248-Elp3i (F), or GV248-Elp4i ( G ). ( H-I ) Hep3B cells were transfected with increasing amount of CMV4-Elp3o (H) or GV248-Elp3i (I). Western blot analysis was performed with anti-PI3K, anti-AKT, anti-p-AKT, anti-Elp3 and anti-Elp4 antibodies. A β-actin western blot is shown as loading control. Results presented as mean ± SD (*P
    Figure Legend Snippet: Elongator activates PI3K/AKT/MMPs signaling pathway. Stably transfected cells were maintained in the absence of serum for 24 h, and then, total RNA or proteins were isolated. Overexpressed Elp3 and Elp4 in HepG2 cells were abbreviated to Elp3o and Elp4o. Elp3i and Elp3i-b were two different clones targeting the same sequence of Elp3 transcript. Elp4i and Elp4i-b were also two different clones targeting the same sequence of Elp4 transcript. ( A ) The mRNA expression of MMP-2 and MMP-9 in stably transfected HepG2 cell lines was examined by qRT-PCR. Related mRNA levels of MMP-2 and MMP-9 were compared between control HepG2 cells and Elp3o or Elp4o cells, and between control HepG2 cells and Elp3i or Elp4i cells. MMP-2 and MMP-9 mRNA levels in control HepG2 cells were set to 1 fold. ( B ) The plasmids for Elp3 overexpression (Elp3o), Elp3 interference (Elp3i), or vector alone (Vector) were transfected into Hep3B cells. The protein expression of Elp3 and Elp4 in transiently transfected Hep3B cells were examined by western blot analysis. GAPDH was shown as a loading control. ( C ) Rescue Experiment. The plasmids for Elp3 overexpression (Elp3o) were transiently transfected into HepG2-Elp3i cells. Cells were subjected to western blot analysis with antibodies against AKT, phosphorylated AKT, MMP-2 and MMP-9. GAPDH was shown as a loading control. ( D-G ) HepG2 cells were transfected with increasing amount of CMV4-Elp3o (D), CMV4-Elp4o (E), GV248-Elp3i (F), or GV248-Elp4i ( G ). ( H-I ) Hep3B cells were transfected with increasing amount of CMV4-Elp3o (H) or GV248-Elp3i (I). Western blot analysis was performed with anti-PI3K, anti-AKT, anti-p-AKT, anti-Elp3 and anti-Elp4 antibodies. A β-actin western blot is shown as loading control. Results presented as mean ± SD (*P

    Techniques Used: Stable Transfection, Transfection, Isolation, Clone Assay, Sequencing, Expressing, Quantitative RT-PCR, Over Expression, Plasmid Preparation, Western Blot

    PI3K/AKT signaling pathway is required for the Elongator-mediated migration and invasion of HCC cells. The plasmids for Elp3 overexpression (Elp3o) and Elp4 overexpression (Elp4o), Elp3 interference (Elp3i) and Elp4 interference (Elp4i) were introduced into stably transfected HepG2 cell lines. Stably transfected cells were maintained with the inhibitor LY294002 (25 μM for 2 hours) when indicated. ( A-B ) Wound-healing assay. Cells were maintained in the absence of serum and allowed to migrate. Wound fields were observed directly after scribing (0 d), and cell migration was followed for 1, 2 and 3 days. Pictures were taken from indicated time phase during 3 days of migration. Representative images were shown in left. Diagrams in right showed the percentage of wound closure at 0, 1, 2 and 3 days. For each experimental condition, the width of starching was set to 100% at time 0 and the width in other time phase expressed relative to that. ( C-D ) Transwell assay. Migration through membrane (C) and invasion through matrigel (D) were carried out with FBS (10%) as achemoattractant in transwell assay. Cells on the lower surface of the chamber were stained by crystal violet after transfection. Representative images of migrating cells or invading cells after staining were taken either in the absence or presence of the inhibitor LY294002. Histograms show the number of migrating cells and invading cells in right. ( E ) Total RNA was extracted and qRT-PCR was performed with primer pairs to amplify Elp3 and Elp4. The relative mRNA levels of Elp3 or Elp4 were examined. Elp3 and Elp4 mRNA levels in control HepG2 cells were set to 1 fold. ( F ) Cell lysates were extracted and subjected to western blot analysis with antibodies as indicated. The protein expressions of Elp3 and Elp4 were compared between inhibitor treatment and untreatment. ( G-H ) Total cell lysates of cells were probed for the expression of Elp3 (G) or Elp4 (H) levels normalized with β-actin by western blot analysis. Western blot was performed with specific antibodies against PI3K, AKT, phosphorylated AKT, MMP-2 and MMP-9.The densitometric analysis for the western blot was shown at the bottom. ( I ) The mRNA expression of MMP-2 and MMP-9 were compared between before and after inhibitor treatment in each group of cells respectively. The graphs represent densitometric results from three independent experiments. Each value represents the mean ± SD (*P
    Figure Legend Snippet: PI3K/AKT signaling pathway is required for the Elongator-mediated migration and invasion of HCC cells. The plasmids for Elp3 overexpression (Elp3o) and Elp4 overexpression (Elp4o), Elp3 interference (Elp3i) and Elp4 interference (Elp4i) were introduced into stably transfected HepG2 cell lines. Stably transfected cells were maintained with the inhibitor LY294002 (25 μM for 2 hours) when indicated. ( A-B ) Wound-healing assay. Cells were maintained in the absence of serum and allowed to migrate. Wound fields were observed directly after scribing (0 d), and cell migration was followed for 1, 2 and 3 days. Pictures were taken from indicated time phase during 3 days of migration. Representative images were shown in left. Diagrams in right showed the percentage of wound closure at 0, 1, 2 and 3 days. For each experimental condition, the width of starching was set to 100% at time 0 and the width in other time phase expressed relative to that. ( C-D ) Transwell assay. Migration through membrane (C) and invasion through matrigel (D) were carried out with FBS (10%) as achemoattractant in transwell assay. Cells on the lower surface of the chamber were stained by crystal violet after transfection. Representative images of migrating cells or invading cells after staining were taken either in the absence or presence of the inhibitor LY294002. Histograms show the number of migrating cells and invading cells in right. ( E ) Total RNA was extracted and qRT-PCR was performed with primer pairs to amplify Elp3 and Elp4. The relative mRNA levels of Elp3 or Elp4 were examined. Elp3 and Elp4 mRNA levels in control HepG2 cells were set to 1 fold. ( F ) Cell lysates were extracted and subjected to western blot analysis with antibodies as indicated. The protein expressions of Elp3 and Elp4 were compared between inhibitor treatment and untreatment. ( G-H ) Total cell lysates of cells were probed for the expression of Elp3 (G) or Elp4 (H) levels normalized with β-actin by western blot analysis. Western blot was performed with specific antibodies against PI3K, AKT, phosphorylated AKT, MMP-2 and MMP-9.The densitometric analysis for the western blot was shown at the bottom. ( I ) The mRNA expression of MMP-2 and MMP-9 were compared between before and after inhibitor treatment in each group of cells respectively. The graphs represent densitometric results from three independent experiments. Each value represents the mean ± SD (*P

    Techniques Used: Migration, Over Expression, Stable Transfection, Transfection, Wound Healing Assay, Transwell Assay, Staining, Quantitative RT-PCR, Western Blot, Expressing

    Elp3 and Elp4 promote cell migration and invasion of HepG2 cells. The plasmids for Elp3 overexpression (Elp3o) and Elp4 overexpression (Elp4o), Elp3 interference (Elp3i) and Elp4 interference (Elp4i) were introduced into stably transfected HepG2 cell lines (A-D). ( A ) The mRNA expression of Elp3 or Elp4 in stably transfected HepG2 cell lines was examined by qRT-PCR as described in Materials and Methods. The relative mRNA levels of Elp3 or Elp4 was normalized against β-actin and depicted graphically. Elp3 and Elp4 mRNA levels in control HepG2 cells were set to 1 fold. ( B ) The protein expression of Elp3 and Elp4 in stably transfected HepG2 cells were examined by western blot analysis. A β-actin western blot is shown as loading control. ( C ) Wound-healing assay. Wound fields were observed directly after scribing (0 d), and cell migration was followed for 1, 2 and 3 days. Representative images taken at 0 and 3 days were shown up in the left panel. A quantification of the data obtained is illustrated on the right. For each experimental condition, the width of starching was set to 100% at time 0 and the width in other time points expressed relative to that. ( D ) Transwell assay. Cells on the lower surface of the chamber were stained by crystal violet after transfection. Representative images of migrated cells or invaded cells are shown in the left panel. The numbers of migration cells and invasion cells were counted. Data were calculated and presented as a histogram in the right panel. The results are expressed as the mean ± SD of three independent experiments. The figures show the data from a representative experiment performed in triplicates (*P
    Figure Legend Snippet: Elp3 and Elp4 promote cell migration and invasion of HepG2 cells. The plasmids for Elp3 overexpression (Elp3o) and Elp4 overexpression (Elp4o), Elp3 interference (Elp3i) and Elp4 interference (Elp4i) were introduced into stably transfected HepG2 cell lines (A-D). ( A ) The mRNA expression of Elp3 or Elp4 in stably transfected HepG2 cell lines was examined by qRT-PCR as described in Materials and Methods. The relative mRNA levels of Elp3 or Elp4 was normalized against β-actin and depicted graphically. Elp3 and Elp4 mRNA levels in control HepG2 cells were set to 1 fold. ( B ) The protein expression of Elp3 and Elp4 in stably transfected HepG2 cells were examined by western blot analysis. A β-actin western blot is shown as loading control. ( C ) Wound-healing assay. Wound fields were observed directly after scribing (0 d), and cell migration was followed for 1, 2 and 3 days. Representative images taken at 0 and 3 days were shown up in the left panel. A quantification of the data obtained is illustrated on the right. For each experimental condition, the width of starching was set to 100% at time 0 and the width in other time points expressed relative to that. ( D ) Transwell assay. Cells on the lower surface of the chamber were stained by crystal violet after transfection. Representative images of migrated cells or invaded cells are shown in the left panel. The numbers of migration cells and invasion cells were counted. Data were calculated and presented as a histogram in the right panel. The results are expressed as the mean ± SD of three independent experiments. The figures show the data from a representative experiment performed in triplicates (*P

    Techniques Used: Migration, Over Expression, Stable Transfection, Transfection, Expressing, Quantitative RT-PCR, Western Blot, Wound Healing Assay, Transwell Assay, Staining

    53) Product Images from "Overexpression of miR-21-5p promotes proliferation and invasion of colon adenocarcinoma cells through targeting CHL1"

    Article Title: Overexpression of miR-21-5p promotes proliferation and invasion of colon adenocarcinoma cells through targeting CHL1

    Journal: Molecular Medicine

    doi: 10.1186/s10020-018-0034-5

    MiR-21-5p highly expressed in COAD tissues and cells. a The heat map showed that miR-21-5p was high expressed in COAD tissues compared with normal tissues. b The relative expression of miR-21-5p in cancer tissues was much higher than that in adjacent tissues detected by qRT-PCR. *** P
    Figure Legend Snippet: MiR-21-5p highly expressed in COAD tissues and cells. a The heat map showed that miR-21-5p was high expressed in COAD tissues compared with normal tissues. b The relative expression of miR-21-5p in cancer tissues was much higher than that in adjacent tissues detected by qRT-PCR. *** P

    Techniques Used: Expressing, Quantitative RT-PCR

    The transfection of miR-21-5p mimics and miR-21-5p inhibitor. a The result of qRT-PCR showed that miR-21-5p expression in miR-21-5p mimics was much higher than that in NC mimics. *** P
    Figure Legend Snippet: The transfection of miR-21-5p mimics and miR-21-5p inhibitor. a The result of qRT-PCR showed that miR-21-5p expression in miR-21-5p mimics was much higher than that in NC mimics. *** P

    Techniques Used: Transfection, Quantitative RT-PCR, Expressing

    54) Product Images from "Aberrant Expression of miR-592 Is Associated with Prognosis and Progression of Renal Cell Carcinoma"

    Article Title: Aberrant Expression of miR-592 Is Associated with Prognosis and Progression of Renal Cell Carcinoma

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S227834

    Relative miR-592 expression in RCC tissues and cell lines. ( A ). The qRT-PCR analysis showed the expression of miR-592 in the RCC tissues and the adjacent normal tissues. Differences were analyzed using 2-tailed Student’s t-test. ( B ). qRT-PCR analysis of miR-592 in RCC cancer cell lines and normal renal epithelial HK-2 cells. Differences were analyzed using one-way ANOVA followed by Tukey’s post-hoc test. *** P
    Figure Legend Snippet: Relative miR-592 expression in RCC tissues and cell lines. ( A ). The qRT-PCR analysis showed the expression of miR-592 in the RCC tissues and the adjacent normal tissues. Differences were analyzed using 2-tailed Student’s t-test. ( B ). qRT-PCR analysis of miR-592 in RCC cancer cell lines and normal renal epithelial HK-2 cells. Differences were analyzed using one-way ANOVA followed by Tukey’s post-hoc test. *** P

    Techniques Used: Expressing, Quantitative RT-PCR

    Effects of miR-592 on the proliferation of 786-O and Caki-1 cells. Differences were analyzed using one-way ANOVA followed by Tukey’s post-hoc test. ( A ). miR-592 mimics, miR-592 inhibitors or NCs were transfected into 786-O and Caki-1 cells, and transfection efficiency was measured using qRT-PCR. ( B ). Cell proliferation was measured by CCK-8 assays. * P
    Figure Legend Snippet: Effects of miR-592 on the proliferation of 786-O and Caki-1 cells. Differences were analyzed using one-way ANOVA followed by Tukey’s post-hoc test. ( A ). miR-592 mimics, miR-592 inhibitors or NCs were transfected into 786-O and Caki-1 cells, and transfection efficiency was measured using qRT-PCR. ( B ). Cell proliferation was measured by CCK-8 assays. * P

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

    Identification of SPRY2 as a direct target of miR-592 in RCC cells. ( A ) Sequence alignment of miR-592 and the 3ʹ-UTR of SPRY2. ( B ) The qRT-PCR analysis was conducted to detect the SPRY2 mRNA in Caki-1 cells transfected with miR-592 mimic, mimic NC, miR-592 inhibitor, or inhibitor NC. Differences were analyzed using one-way ANOVA followed by Tukey’s post-hoc test. ( C ) Luciferase reporter assay in Caki-1 cells that were co-transfected with miR-592 mimics, mimic NC, miR-592 inhibitor, or inhibitor NC and Wt-type SPRY2 3ʹ-UTR vector of Mut-type vector. Differences were analyzed using one-way ANOVA followed by Tukey’s post-hoc test. * P
    Figure Legend Snippet: Identification of SPRY2 as a direct target of miR-592 in RCC cells. ( A ) Sequence alignment of miR-592 and the 3ʹ-UTR of SPRY2. ( B ) The qRT-PCR analysis was conducted to detect the SPRY2 mRNA in Caki-1 cells transfected with miR-592 mimic, mimic NC, miR-592 inhibitor, or inhibitor NC. Differences were analyzed using one-way ANOVA followed by Tukey’s post-hoc test. ( C ) Luciferase reporter assay in Caki-1 cells that were co-transfected with miR-592 mimics, mimic NC, miR-592 inhibitor, or inhibitor NC and Wt-type SPRY2 3ʹ-UTR vector of Mut-type vector. Differences were analyzed using one-way ANOVA followed by Tukey’s post-hoc test. * P

    Techniques Used: Sequencing, Quantitative RT-PCR, Transfection, Luciferase, Reporter Assay, Plasmid Preparation

    55) Product Images from "Long noncoding RNA OR3A4 promotes the proliferation and invasion of osteosarcoma cells by sponging miR-1227-5p"

    Article Title: Long noncoding RNA OR3A4 promotes the proliferation and invasion of osteosarcoma cells by sponging miR-1227-5p

    Journal: Journal of Bone Oncology

    doi: 10.1016/j.jbo.2020.100278

    Downregulation of OR3A4 inhibited the proliferation and invasion of osteosarcoma cells. (A) qRT-PCR was used to detect the expression of OR3A4 in the MG-63 cell line transfected with si-OR3A4. (B) MTT assay was performed to analyse the effect of OR3A4 on MG-63 cell proliferation. (C) Transwell assay was conducted to determine the effect of OR3A4 on MG-63 cell invasion ** p
    Figure Legend Snippet: Downregulation of OR3A4 inhibited the proliferation and invasion of osteosarcoma cells. (A) qRT-PCR was used to detect the expression of OR3A4 in the MG-63 cell line transfected with si-OR3A4. (B) MTT assay was performed to analyse the effect of OR3A4 on MG-63 cell proliferation. (C) Transwell assay was conducted to determine the effect of OR3A4 on MG-63 cell invasion ** p

    Techniques Used: Quantitative RT-PCR, Expressing, Transfection, MTT Assay, Transwell Assay

    MiR-1227-5p was the potential target microRNA of OR3A4, which was predicted and validated by bioinformatics analysis and luciferase reporter assay. (A) A flow chart to screen the miRNAs based on the diagrams. (B) The complementary binding site of OR3A4 and miR-1227-5p was predicted by bioinformatics. (C) qRT-PCR was performed to observe the expression of miR-1227-5p in MG-63 cells transfected with si-OR3A4. (D) Luciferase reporter assay was used to confirm the predicted binding. (E) The expression of miR-1227-5p in clinical stages I/II and III/IV of osteosarcoma was detected by qRT-PCR. ** p
    Figure Legend Snippet: MiR-1227-5p was the potential target microRNA of OR3A4, which was predicted and validated by bioinformatics analysis and luciferase reporter assay. (A) A flow chart to screen the miRNAs based on the diagrams. (B) The complementary binding site of OR3A4 and miR-1227-5p was predicted by bioinformatics. (C) qRT-PCR was performed to observe the expression of miR-1227-5p in MG-63 cells transfected with si-OR3A4. (D) Luciferase reporter assay was used to confirm the predicted binding. (E) The expression of miR-1227-5p in clinical stages I/II and III/IV of osteosarcoma was detected by qRT-PCR. ** p

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

    56) Product Images from "A MicroRNA-Mediated Insulin Signaling Pathway Regulates the Toxicity of Multi-Walled Carbon Nanotubes in Nematode Caenorhabditis elegans"

    Article Title: A MicroRNA-Mediated Insulin Signaling Pathway Regulates the Toxicity of Multi-Walled Carbon Nanotubes in Nematode Caenorhabditis elegans

    Journal: Scientific Reports

    doi: 10.1038/srep23234

    Dysregulated mRNAs induced by MWCNTs exposure. ( a ) Heatmap showing the expression of mRNAs obtained from control and MWCNTs-exposed nematodes. Relatively low expression levels are represented as blue, and relatively high expression levels are represented in red. ( b ) Scatter diagram of relationship between mRNA coverage of the control group and the MWCNTs exposure group. ( c ) qRT-PCR analysis of the expression expressions of some genes encoding insulin signaling pathway in nematodes exposed to MWCNTs. ( d ) MWCNTs exposure influenced the nuclear translocation of DAF-16::GFP in nematodes. Scale bar, 150 μm. MWCNTs (1 mg/L) exposure was performed from L1-larvae to young adult. Bars represent means ± S.E.M. ** P
    Figure Legend Snippet: Dysregulated mRNAs induced by MWCNTs exposure. ( a ) Heatmap showing the expression of mRNAs obtained from control and MWCNTs-exposed nematodes. Relatively low expression levels are represented as blue, and relatively high expression levels are represented in red. ( b ) Scatter diagram of relationship between mRNA coverage of the control group and the MWCNTs exposure group. ( c ) qRT-PCR analysis of the expression expressions of some genes encoding insulin signaling pathway in nematodes exposed to MWCNTs. ( d ) MWCNTs exposure influenced the nuclear translocation of DAF-16::GFP in nematodes. Scale bar, 150 μm. MWCNTs (1 mg/L) exposure was performed from L1-larvae to young adult. Bars represent means ± S.E.M. ** P

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

    57) Product Images from "Long non-coding RNA HOTAIR is associated with human cervical cancer progression"

    Article Title: Long non-coding RNA HOTAIR is associated with human cervical cancer progression

    Journal: International Journal of Oncology

    doi: 10.3892/ijo.2014.2758

    HOTAIR increases VEGF and MMP-9 expression in cervical cancer cells. Protein lysates were obtained from siHOTAIR and siNC-transfected HeLa cells 48 h post-transfection. (A) VEGF and (B) MMP-9 expression were analyzed by western blotting. (C) VEGF and MMP-9 levels were analyzed by western blotting in HOTAIR overexpression SiHa cells. Band intensities were quantitated, and VEGF and MMP-9 expression were normalized to that of β-actin. (D) VEGF and MMP-9 levels were determined by qRT-PCR in low groups and high HOTAIR expression groups of cervical cancer tissues. Each assay was performed in triplicate. Data are mean ± SD. * P
    Figure Legend Snippet: HOTAIR increases VEGF and MMP-9 expression in cervical cancer cells. Protein lysates were obtained from siHOTAIR and siNC-transfected HeLa cells 48 h post-transfection. (A) VEGF and (B) MMP-9 expression were analyzed by western blotting. (C) VEGF and MMP-9 levels were analyzed by western blotting in HOTAIR overexpression SiHa cells. Band intensities were quantitated, and VEGF and MMP-9 expression were normalized to that of β-actin. (D) VEGF and MMP-9 levels were determined by qRT-PCR in low groups and high HOTAIR expression groups of cervical cancer tissues. Each assay was performed in triplicate. Data are mean ± SD. * P

    Techniques Used: Expressing, Transfection, Western Blot, Over Expression, Quantitative RT-PCR

    Relative HOTAIR expression and its clinical significance. (A) HOTAIR expression was significantly higher in cervical cancer tissues (n=111) than in non-cancerous tissues (n=40). Relative HOTAIR expression was determined using qRT-PCR with U6 as an internal control. Data are expressed as mean ± SD. * P
    Figure Legend Snippet: Relative HOTAIR expression and its clinical significance. (A) HOTAIR expression was significantly higher in cervical cancer tissues (n=111) than in non-cancerous tissues (n=40). Relative HOTAIR expression was determined using qRT-PCR with U6 as an internal control. Data are expressed as mean ± SD. * P

    Techniques Used: Expressing, Quantitative RT-PCR

    Expression of HOTAIR knockdown on the EMT-related genes in HeLa cells. (A) HeLa cells were transfected with HOTAIR -specific siRNA and siNC for 48 h. E-cadherin, β-catenin, Vimentin, Snail and Twist expression were analyzed by (A) qRT-PCR and (B) western blotting. Each assay was performed in triplicate. Data are mean ± SD. * P
    Figure Legend Snippet: Expression of HOTAIR knockdown on the EMT-related genes in HeLa cells. (A) HeLa cells were transfected with HOTAIR -specific siRNA and siNC for 48 h. E-cadherin, β-catenin, Vimentin, Snail and Twist expression were analyzed by (A) qRT-PCR and (B) western blotting. Each assay was performed in triplicate. Data are mean ± SD. * P

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

    Knockdown of HOTAIR inhibits the cell proliferation of cervical cancer cells. (A) Expression of HOTAIR in cervical cancer cells. HOTAIR expression was evaluated using qRT-PCR with U6 as an internal control. (B) Cells were transfected with HOTAIR -specific siRNA and negative control siRNA (siNC), and knockdown efficiency was determined by qRT-PCR analysis. (C) Knockdown of HOTAIR decreases cell proliferation in HeLa, SiHa and CasKi cells. The proliferation of cervical cancer cells transfected with siHOTAIR and negative control siRNA (siNC) was determined using the CCK-8 assay. Bars indicate mean ± SD of three independent experiments performed in triplicate. * P
    Figure Legend Snippet: Knockdown of HOTAIR inhibits the cell proliferation of cervical cancer cells. (A) Expression of HOTAIR in cervical cancer cells. HOTAIR expression was evaluated using qRT-PCR with U6 as an internal control. (B) Cells were transfected with HOTAIR -specific siRNA and negative control siRNA (siNC), and knockdown efficiency was determined by qRT-PCR analysis. (C) Knockdown of HOTAIR decreases cell proliferation in HeLa, SiHa and CasKi cells. The proliferation of cervical cancer cells transfected with siHOTAIR and negative control siRNA (siNC) was determined using the CCK-8 assay. Bars indicate mean ± SD of three independent experiments performed in triplicate. * P

    Techniques Used: Expressing, Quantitative RT-PCR, Transfection, Negative Control, CCK-8 Assay

    HOTAIR knockdown inhibits the migration and invasion of cervical cancer cells. (A) Wound healing assay was used to determine migration in siHOTAIR-transfected HeLa SiHa and CasKi cells (magnification, ×200). (B) Matrigel invasion assay was used to determine invasion after 48 h in HeLa cells. (C) Overexpression of HOTAIR in SiHa cells analyzed by qRT-PCR. (D) Cell invasion was evaluated using Matrigel invasion chamber. Overexpression of HOTAIR in SiHa cells increased the invasive capacity after 48 h. Each assay was performed in triplicate. Data are mean ± SD. * P
    Figure Legend Snippet: HOTAIR knockdown inhibits the migration and invasion of cervical cancer cells. (A) Wound healing assay was used to determine migration in siHOTAIR-transfected HeLa SiHa and CasKi cells (magnification, ×200). (B) Matrigel invasion assay was used to determine invasion after 48 h in HeLa cells. (C) Overexpression of HOTAIR in SiHa cells analyzed by qRT-PCR. (D) Cell invasion was evaluated using Matrigel invasion chamber. Overexpression of HOTAIR in SiHa cells increased the invasive capacity after 48 h. Each assay was performed in triplicate. Data are mean ± SD. * P

    Techniques Used: Migration, Wound Healing Assay, Transfection, Invasion Assay, Over Expression, Quantitative RT-PCR

    58) Product Images from "Derivate Isocorydine (d-ICD) Suppresses Migration and Invasion of Hepatocellular Carcinoma Cell by Downregulating ITGA1 Expression"

    Article Title: Derivate Isocorydine (d-ICD) Suppresses Migration and Invasion of Hepatocellular Carcinoma Cell by Downregulating ITGA1 Expression

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms18030514

    E2F1 transcriptional upregulates ITGA1 expression and d-ICD inhibits E2F1 expression in HCC cells. ( A ) Identification of potential E2F1 binding sites (double arrow marked) in the ITGA1 promoter (arrow direction indicates sequence from 5′ to 3′) according to four TF-binding prediction websites (QIAGEN, ALGGEN PROMO, TFBIND, GENE REGULATION), and the mutant sites are marked (arrow direction indicates transcriptional direction of the downstreem gene); ( B ) Dual luciferase reporter gene studies were performed to detect the ITGA1 promoter and its truncated and mutant construct activity in 293T cells; ( C ) Binding of E2F1 to the ITGA1 promoter in HCC cells was analyzed by chromatin immunoprecipitation. qRT-PCR was used to analyze the ITGA1 promoter, and agarose gel electrophoresis was used to analyze the crosslinking status; ( D ) qRT-PCR and western blotting analyzed E2F1 and ITGA1 expression in HCC cells with transiently transfected E2F1 plasmid; ( E ) Western blotting analyses of E2F1 expression in HCC cells treated with d-ICD at different time points. (* p
    Figure Legend Snippet: E2F1 transcriptional upregulates ITGA1 expression and d-ICD inhibits E2F1 expression in HCC cells. ( A ) Identification of potential E2F1 binding sites (double arrow marked) in the ITGA1 promoter (arrow direction indicates sequence from 5′ to 3′) according to four TF-binding prediction websites (QIAGEN, ALGGEN PROMO, TFBIND, GENE REGULATION), and the mutant sites are marked (arrow direction indicates transcriptional direction of the downstreem gene); ( B ) Dual luciferase reporter gene studies were performed to detect the ITGA1 promoter and its truncated and mutant construct activity in 293T cells; ( C ) Binding of E2F1 to the ITGA1 promoter in HCC cells was analyzed by chromatin immunoprecipitation. qRT-PCR was used to analyze the ITGA1 promoter, and agarose gel electrophoresis was used to analyze the crosslinking status; ( D ) qRT-PCR and western blotting analyzed E2F1 and ITGA1 expression in HCC cells with transiently transfected E2F1 plasmid; ( E ) Western blotting analyses of E2F1 expression in HCC cells treated with d-ICD at different time points. (* p

    Techniques Used: Expressing, Binding Assay, Sequencing, Mutagenesis, Luciferase, Construct, Activity Assay, Chromatin Immunoprecipitation, Quantitative RT-PCR, Agarose Gel Electrophoresis, Western Blot, Transfection, Plasmid Preparation

    Silencing ITGA1 expression inhibits HCC cell migration and invasion in vitro. ( A ) qRT-PCR and western blot analyzed the silencing efficiency of ITGA1 expression; ( B ) The in vitro migration ability of SMMC-7721, Huh7 and HCC-LY5 cells stably transfected with ITGA1 ShRNA or ShNC or MOCK cells were assessed using wound-healing assay (scale bar stand for 200 μm); ( C ) The in vitro invasion ability of those ITGA1 -silencing HCC cells were assessed by trans-well assay (scale bar stand for 100 μm). (“ns” indicates no statistical significance, * p
    Figure Legend Snippet: Silencing ITGA1 expression inhibits HCC cell migration and invasion in vitro. ( A ) qRT-PCR and western blot analyzed the silencing efficiency of ITGA1 expression; ( B ) The in vitro migration ability of SMMC-7721, Huh7 and HCC-LY5 cells stably transfected with ITGA1 ShRNA or ShNC or MOCK cells were assessed using wound-healing assay (scale bar stand for 200 μm); ( C ) The in vitro invasion ability of those ITGA1 -silencing HCC cells were assessed by trans-well assay (scale bar stand for 100 μm). (“ns” indicates no statistical significance, * p

    Techniques Used: Expressing, Migration, In Vitro, Quantitative RT-PCR, Western Blot, Stable Transfection, Transfection, shRNA, Wound Healing Assay

    59) Product Images from "Long noncoding RNA NBAT-1 suppresses tumorigenesis and predicts favorable prognosis in ovarian cancer"

    Article Title: Long noncoding RNA NBAT-1 suppresses tumorigenesis and predicts favorable prognosis in ovarian cancer

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S124645

    lncRNA NBAT-1 expression in GEO and human OC tissues. Notes: ( A ) lncRNA NBAT-1 was consistently downregulated in OC tissue compared to the normal tissue in both GSE18520 and GSE38666. ( B ) Relative expression of NBAT-1 in EOC tissue (n=57) and normal tissue (n=29) was examined by RT-PCR and normalized to GAPDH expression. ( C ) lncRNA NBAT-1 expression was classified into two groups, according to the median expression level in OC tissues. ( D ) The correlation between NBAT-1 expression and prognosis. Five-year overall survival was analyzed by the Kaplan–Meier survival curve. Abbreviations: EOC, epithelial ovarian cancer; GEO, Gene Expression Omnibus; lncRNA, long noncoding RNA; OC, ovarian cancer; RT-PCR, real-time polymerase chain reaction.
    Figure Legend Snippet: lncRNA NBAT-1 expression in GEO and human OC tissues. Notes: ( A ) lncRNA NBAT-1 was consistently downregulated in OC tissue compared to the normal tissue in both GSE18520 and GSE38666. ( B ) Relative expression of NBAT-1 in EOC tissue (n=57) and normal tissue (n=29) was examined by RT-PCR and normalized to GAPDH expression. ( C ) lncRNA NBAT-1 expression was classified into two groups, according to the median expression level in OC tissues. ( D ) The correlation between NBAT-1 expression and prognosis. Five-year overall survival was analyzed by the Kaplan–Meier survival curve. Abbreviations: EOC, epithelial ovarian cancer; GEO, Gene Expression Omnibus; lncRNA, long noncoding RNA; OC, ovarian cancer; RT-PCR, real-time polymerase chain reaction.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

    NBAT-1 suppresses tumor growth in vivo. Notes: ( A and B ) Empty vector and pcDNA-NBAT-1 were transfected into OVCAR-3 cells, which were injected in the nude mice (n=6). Tumor volumes were calculated after injection every 3 days. ( C ) Tumor weight are represented as mean of tumor weights ± SEM. qRT-PCR was performed to detect the average expression of NBAT-1 in xenograft tumors (n=6). ( D ) The tumor sections were under hematoxylin-eosin staining and immunohistochemistry staining using antibodies against Ki-67. ***( P
    Figure Legend Snippet: NBAT-1 suppresses tumor growth in vivo. Notes: ( A and B ) Empty vector and pcDNA-NBAT-1 were transfected into OVCAR-3 cells, which were injected in the nude mice (n=6). Tumor volumes were calculated after injection every 3 days. ( C ) Tumor weight are represented as mean of tumor weights ± SEM. qRT-PCR was performed to detect the average expression of NBAT-1 in xenograft tumors (n=6). ( D ) The tumor sections were under hematoxylin-eosin staining and immunohistochemistry staining using antibodies against Ki-67. ***( P

    Techniques Used: In Vivo, Plasmid Preparation, Transfection, Injection, Mouse Assay, Quantitative RT-PCR, Expressing, Staining, Immunohistochemistry

    Overexpressed NBAT-1 inhibits OC cell proliferation in vitro. Notes: ( A ) Expression of lncRNA NBAT-1 in five EOC cell lines was determined by qRT-PCR. ( B ) Overexpressed NBAT-1 efficiency was determined by qRT-PCR in TOV112D and OVCAR-3 cells. ( C and D ) Overexpressed NBAT-1 in TOV112D and OVCAR-3 cells significantly reduced their proliferative capacities, as determined by a cell number counting assay and colony formation assay. ***( P
    Figure Legend Snippet: Overexpressed NBAT-1 inhibits OC cell proliferation in vitro. Notes: ( A ) Expression of lncRNA NBAT-1 in five EOC cell lines was determined by qRT-PCR. ( B ) Overexpressed NBAT-1 efficiency was determined by qRT-PCR in TOV112D and OVCAR-3 cells. ( C and D ) Overexpressed NBAT-1 in TOV112D and OVCAR-3 cells significantly reduced their proliferative capacities, as determined by a cell number counting assay and colony formation assay. ***( P

    Techniques Used: In Vitro, Expressing, Quantitative RT-PCR, Colony Assay

    60) Product Images from "Screening differential circular RNA expression profiles reveal that hsa_circ_0128298 is a biomarker in the diagnosis and prognosis of hepatocellular carcinoma"

    Article Title: Screening differential circular RNA expression profiles reveal that hsa_circ_0128298 is a biomarker in the diagnosis and prognosis of hepatocellular carcinoma

    Journal: Cancer Management and Research

    doi: 10.2147/CMAR.S166740

    The detailed experimental process. Notes: circRNA expression profiles were screened in three paired human HCC and paratumorous tissues. Targeted circRNAs were verified by qRT-PCR via initial verification in 30 pairs of samples and extended verification in 78 pairs of samples. Abbreviations: circRNAs, circular RNAs; HCC, hepatocellular carcinoma; qRT-PCR, quantitative real-time polymerase chain reaction.
    Figure Legend Snippet: The detailed experimental process. Notes: circRNA expression profiles were screened in three paired human HCC and paratumorous tissues. Targeted circRNAs were verified by qRT-PCR via initial verification in 30 pairs of samples and extended verification in 78 pairs of samples. Abbreviations: circRNAs, circular RNAs; HCC, hepatocellular carcinoma; qRT-PCR, quantitative real-time polymerase chain reaction.

    Techniques Used: Expressing, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

    61) Product Images from "TNF-α-Induced SOX5 Upregulation Is Involved in the Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells Through KLF4 Signal Pathway"

    Article Title: TNF-α-Induced SOX5 Upregulation Is Involved in the Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells Through KLF4 Signal Pathway

    Journal: Molecules and Cells

    doi: 10.14348/molcells.2018.2359

    SOX5 mediated osteogenic differentiation of hMSCs through KLF4 signal pathway The KLF4 mRNA and protein expression were examined by qRT-PCR (A, C, and I) and western blotting (B, D, and H). ALP activity was detected by a commercial Alkaline Phosphatase Detection Kit (E, J). The genes expression of osteogenic marker including Collagen I, Runx2 and Osterix were examined by Western blotting (F, K) and qRT-PCR (G, L). pcSOX5+siKLF4: cells were transfected with recombinant plasmid pcDNA3.1/SOX5 (pcSOX5) and KLF4 siRNA, and then cultured with OM. TNF-α+siKLF4: cells were transfected with KLF4 siRNA and then cultured with OM containing TNF-α. Data are expressed as the mean ± SD, n = 3. *P
    Figure Legend Snippet: SOX5 mediated osteogenic differentiation of hMSCs through KLF4 signal pathway The KLF4 mRNA and protein expression were examined by qRT-PCR (A, C, and I) and western blotting (B, D, and H). ALP activity was detected by a commercial Alkaline Phosphatase Detection Kit (E, J). The genes expression of osteogenic marker including Collagen I, Runx2 and Osterix were examined by Western blotting (F, K) and qRT-PCR (G, L). pcSOX5+siKLF4: cells were transfected with recombinant plasmid pcDNA3.1/SOX5 (pcSOX5) and KLF4 siRNA, and then cultured with OM. TNF-α+siKLF4: cells were transfected with KLF4 siRNA and then cultured with OM containing TNF-α. Data are expressed as the mean ± SD, n = 3. *P

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, ALP Assay, Activity Assay, Marker, Transfection, Recombinant, Plasmid Preparation, Cell Culture

    SOX5 overexpression inhibited osteogenic differentiation of hMSCs SOX5 mRNA and protein expression were examined by qRTPCR (A) and western blotting (B) in cells cultured with OM for 0 days (control) and 14 days (OM). Cells were transfected with recombinant plasmid pcDNA3.1/SOX5 (pcSOX5) and its negative control plasmid (pcNC), or SOX5 siRNA (siSOX5) and its negative control (siNC) for 15 h, after which the medium was replaced with OM for 14 days. The efficiency of transfection was evaluated by qRT-PCR (C) and Western blotting (D). ALP activity was detected by a commercial Alkaline Phosphatase Detection Kit (E). The genes expression of osteogenic marker including Collagen I, Runx2 and Osterix were examined by Western blotting (F) and qRT-PCR (G). Data are expressed as the mean ± SD, n = 3. *P
    Figure Legend Snippet: SOX5 overexpression inhibited osteogenic differentiation of hMSCs SOX5 mRNA and protein expression were examined by qRTPCR (A) and western blotting (B) in cells cultured with OM for 0 days (control) and 14 days (OM). Cells were transfected with recombinant plasmid pcDNA3.1/SOX5 (pcSOX5) and its negative control plasmid (pcNC), or SOX5 siRNA (siSOX5) and its negative control (siNC) for 15 h, after which the medium was replaced with OM for 14 days. The efficiency of transfection was evaluated by qRT-PCR (C) and Western blotting (D). ALP activity was detected by a commercial Alkaline Phosphatase Detection Kit (E). The genes expression of osteogenic marker including Collagen I, Runx2 and Osterix were examined by Western blotting (F) and qRT-PCR (G). Data are expressed as the mean ± SD, n = 3. *P

    Techniques Used: Over Expression, Expressing, Western Blot, Cell Culture, Transfection, Recombinant, Plasmid Preparation, Negative Control, Quantitative RT-PCR, ALP Assay, Activity Assay, Marker

    TNF-α induced SOX5 gene expression during osteogenic differentiation of hMSCs The SOX5 mRNA and protein expression were examined by qRT-PCR (A) and western blotting (B). ALP activity was detected by a commercial Alkaline Phosphatase Detection Kit (C). The genes expression of osteogenic marker including Collagen I, Runx2 and Osterix were examined by Western blotting (D) and qRTPCR (E). TNF-α: cells were cultured with OM containing TNF-α; TNF-α+siSOX5: cells were transfected with SOX5 siRNA and then cultured with OM containing TNF-α. Data are expressed as the mean ± SD, n = 3. *P
    Figure Legend Snippet: TNF-α induced SOX5 gene expression during osteogenic differentiation of hMSCs The SOX5 mRNA and protein expression were examined by qRT-PCR (A) and western blotting (B). ALP activity was detected by a commercial Alkaline Phosphatase Detection Kit (C). The genes expression of osteogenic marker including Collagen I, Runx2 and Osterix were examined by Western blotting (D) and qRTPCR (E). TNF-α: cells were cultured with OM containing TNF-α; TNF-α+siSOX5: cells were transfected with SOX5 siRNA and then cultured with OM containing TNF-α. Data are expressed as the mean ± SD, n = 3. *P

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, ALP Assay, Activity Assay, Marker, Cell Culture, Transfection

    SOX5 was up-regulated in hMSCs isolated from bone marrow samples of patients with postmenopausal osteoporosis (PMOP) and healthy premenopausal women (healthy control), the mRNA and protein expression levels of SOX5 were examined by qRT-PCR (A) and Western blotting (B). Data are expressed as the mean ± SD, n=3. **P
    Figure Legend Snippet: SOX5 was up-regulated in hMSCs isolated from bone marrow samples of patients with postmenopausal osteoporosis (PMOP) and healthy premenopausal women (healthy control), the mRNA and protein expression levels of SOX5 were examined by qRT-PCR (A) and Western blotting (B). Data are expressed as the mean ± SD, n=3. **P

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

    62) Product Images from "Reduced C9orf72 gene expression in c9FTD/ALS is caused by histone trimethylation, an epigenetic event detectable in blood"

    Article Title: Reduced C9orf72 gene expression in c9FTD/ALS is caused by histone trimethylation, an epigenetic event detectable in blood

    Journal: Acta Neuropathologica

    doi: 10.1007/s00401-013-1199-1

    C9orf72 mRNA expression is increased and H3K9me3 binding is decreased in C9orf72+ fibroblasts upon 5-AZA treatment. a , b qRT-PCR of RNA obtained from human fibroblasts grown in DMSO or 5-AZA demethylating agent. Both assays targeting transcript variants 1, 2, 3 ( a ) and 2, 3 ( b ) show a significant increase in expression after 5-AZA treatment only in C9orf72 repeat expansion carriers. c qRT-PCR of H19 , an imprinted gene, showing effectiveness of the 5-AZA treatment in C9orf72+ and C9orf72− fibroblasts. Statistical differences were assessed by unpaired Student t test. * p
    Figure Legend Snippet: C9orf72 mRNA expression is increased and H3K9me3 binding is decreased in C9orf72+ fibroblasts upon 5-AZA treatment. a , b qRT-PCR of RNA obtained from human fibroblasts grown in DMSO or 5-AZA demethylating agent. Both assays targeting transcript variants 1, 2, 3 ( a ) and 2, 3 ( b ) show a significant increase in expression after 5-AZA treatment only in C9orf72 repeat expansion carriers. c qRT-PCR of H19 , an imprinted gene, showing effectiveness of the 5-AZA treatment in C9orf72+ and C9orf72− fibroblasts. Statistical differences were assessed by unpaired Student t test. * p

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

    Frontal cortex and cerebellum tissue samples from the C9orf72+ group exhibit a significant decrease in C9orf72 mRNA expression levels. a Schematic representation showing the three known transcript variants of C9orf72 . The star marks the repeat location. b , c qRT-PCR for C9orf72 transcript variants 1, 2, 3 and C9orf72 transcript variants 2, 3 were performed in frontal cortex ( b ) and cerebellum ( c ). Data on graphs were normalized to disease control group (mean value set to 1). Statistical differences were calculated by one-way ANOVA with Tukey post hoc test. * p
    Figure Legend Snippet: Frontal cortex and cerebellum tissue samples from the C9orf72+ group exhibit a significant decrease in C9orf72 mRNA expression levels. a Schematic representation showing the three known transcript variants of C9orf72 . The star marks the repeat location. b , c qRT-PCR for C9orf72 transcript variants 1, 2, 3 and C9orf72 transcript variants 2, 3 were performed in frontal cortex ( b ) and cerebellum ( c ). Data on graphs were normalized to disease control group (mean value set to 1). Statistical differences were calculated by one-way ANOVA with Tukey post hoc test. * p

    Techniques Used: Expressing, Quantitative RT-PCR

    63) Product Images from "Upregulation of long noncoding RNA TUG1 promotes cervical cancer cell proliferation and migration"

    Article Title: Upregulation of long noncoding RNA TUG1 promotes cervical cancer cell proliferation and migration

    Journal: Cancer Medicine

    doi: 10.1002/cam4.994

    TUG 1 is upregulated in cervical cancer tissues and cell lines. (A) The relative expression of TUG 1 was increased in cervical cancer and cervical intraepithelial neoplasia ( CIN ) tissues, as determined by quantitative real‐time polymerase chain reaction ( qRT ‐ PCR ) normalized against unpaired normal samples. (B) Expression of TUG 1 in SiHa, CaSki, HeLa, and C33A cells was measured and normalized against that of unpaired normal samples using qRT ‐ PCR . All experiments were performed in triplicate. Data are presented as means ± SEM . * P
    Figure Legend Snippet: TUG 1 is upregulated in cervical cancer tissues and cell lines. (A) The relative expression of TUG 1 was increased in cervical cancer and cervical intraepithelial neoplasia ( CIN ) tissues, as determined by quantitative real‐time polymerase chain reaction ( qRT ‐ PCR ) normalized against unpaired normal samples. (B) Expression of TUG 1 in SiHa, CaSki, HeLa, and C33A cells was measured and normalized against that of unpaired normal samples using qRT ‐ PCR . All experiments were performed in triplicate. Data are presented as means ± SEM . * P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

    64) Product Images from "circRNA circ‐CCND1 promotes the proliferation of laryngeal squamous cell carcinoma through elevating CCND1 expression via interacting with HuR and miR‐646. circRNA circ‐CCND1 promotes the proliferation of laryngeal squamous cell carcinoma through elevating CCND1 expression via interacting with HuR and miR‐646"

    Article Title: circRNA circ‐CCND1 promotes the proliferation of laryngeal squamous cell carcinoma through elevating CCND1 expression via interacting with HuR and miR‐646. circRNA circ‐CCND1 promotes the proliferation of laryngeal squamous cell carcinoma through elevating CCND1 expression via interacting with HuR and miR‐646

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.14925

    circ‐CCND1 interacts with HuR to increase CCND1 mRNA stability. A, qRT‐PCR analysis for circ‐CCND1 expression in the cytoplasmic and nuclear fractions. B, RNA pull‐down assay in control and circ‐CCND1‐silenced Hep‐2 and TU212 cells, followed by detection of HuR expression using Western blot assay. C, RIP assay in control and circ‐CCND1‐silenced Hep‐2 and TU212 cells, followed by qRT‐PCR analysis for circ‐CCND1 expression. D, The interaction profile between HuR and circ‐CCND1 predicted by online catRAPID algorithm. E, RNA pull‐down assay in Hep‐2 and TU212 cells with indicated wild‐type or mutated probe, followed by Western blot analysis for HuR protein expression. F, RIP assay in control and circ‐CCND1‐silenced Hep‐2 and TU212 cells with HuR or IgG antibody, followed by qRT‐PCR analysis for CCND1 3′‐UTR enrichment. G, qRT‐PCR analysis for CCND1 mRNA expression in control and circ‐CCND1‐silenced Hep‐2 and TU212 cells transfected with HuR expression vector. * P
    Figure Legend Snippet: circ‐CCND1 interacts with HuR to increase CCND1 mRNA stability. A, qRT‐PCR analysis for circ‐CCND1 expression in the cytoplasmic and nuclear fractions. B, RNA pull‐down assay in control and circ‐CCND1‐silenced Hep‐2 and TU212 cells, followed by detection of HuR expression using Western blot assay. C, RIP assay in control and circ‐CCND1‐silenced Hep‐2 and TU212 cells, followed by qRT‐PCR analysis for circ‐CCND1 expression. D, The interaction profile between HuR and circ‐CCND1 predicted by online catRAPID algorithm. E, RNA pull‐down assay in Hep‐2 and TU212 cells with indicated wild‐type or mutated probe, followed by Western blot analysis for HuR protein expression. F, RIP assay in control and circ‐CCND1‐silenced Hep‐2 and TU212 cells with HuR or IgG antibody, followed by qRT‐PCR analysis for CCND1 3′‐UTR enrichment. G, qRT‐PCR analysis for CCND1 mRNA expression in control and circ‐CCND1‐silenced Hep‐2 and TU212 cells transfected with HuR expression vector. * P

    Techniques Used: Quantitative RT-PCR, Expressing, Pull Down Assay, Western Blot, Transfection, Plasmid Preparation

    circ‐CCND1 functions as a sponge for miR‐646. A, The predicted miRNAs bound by circ‐CCND1 using the online CircInteractome tool. B, The specific binding site between circ‐CCND1 and miR‐646. C and D, The luciferase reporter assay in Hep‐2 and TU212 cells cotransfected with control or miR‐646 mimics and wild‐type or mutant circ‐CCND1/CCND1 luciferase vector. E. RNA pull‐down assay in Hep‐2 and TU212 cells using control or circ‐CCND1 probe, followed by qRT‐PCR analysis for miR‐646 expression. F, RNA pull‐down assay in Hep‐2 and TU212 cells transfected with wild‐type or mutant miR‐646 mimics, followed by qRT‐PCR analysis for the enrichment of circ‐CCND1 and CCND1 3′‐UTR. G, qRT‐PCR analysis for miR‐646 expression in control and circ‐CCND1‐silenced Hep‐2 and TU212 cells. H. qRT‐PCR analysis for CCND1 mRNA expression in circ‐CCND1‐silenced Hep‐2 and TU212 cells transfected with miR‐646 inhibitors or combined with HuR expression vector. I, The proliferative capabilities of circ‐CCND1‐silenced Hep‐2 and TU212 cells after transfection with miR‐646 inhibitors or combined with HuR expression vector. J, The schematic illustration of the proposed model depicting the crucial role of circ‐CCND1 in LSCC growth through regulating CCND1 mRNA stability via interaction with HuR and miR‐646. * P
    Figure Legend Snippet: circ‐CCND1 functions as a sponge for miR‐646. A, The predicted miRNAs bound by circ‐CCND1 using the online CircInteractome tool. B, The specific binding site between circ‐CCND1 and miR‐646. C and D, The luciferase reporter assay in Hep‐2 and TU212 cells cotransfected with control or miR‐646 mimics and wild‐type or mutant circ‐CCND1/CCND1 luciferase vector. E. RNA pull‐down assay in Hep‐2 and TU212 cells using control or circ‐CCND1 probe, followed by qRT‐PCR analysis for miR‐646 expression. F, RNA pull‐down assay in Hep‐2 and TU212 cells transfected with wild‐type or mutant miR‐646 mimics, followed by qRT‐PCR analysis for the enrichment of circ‐CCND1 and CCND1 3′‐UTR. G, qRT‐PCR analysis for miR‐646 expression in control and circ‐CCND1‐silenced Hep‐2 and TU212 cells. H. qRT‐PCR analysis for CCND1 mRNA expression in circ‐CCND1‐silenced Hep‐2 and TU212 cells transfected with miR‐646 inhibitors or combined with HuR expression vector. I, The proliferative capabilities of circ‐CCND1‐silenced Hep‐2 and TU212 cells after transfection with miR‐646 inhibitors or combined with HuR expression vector. J, The schematic illustration of the proposed model depicting the crucial role of circ‐CCND1 in LSCC growth through regulating CCND1 mRNA stability via interaction with HuR and miR‐646. * P

    Techniques Used: Binding Assay, Luciferase, Reporter Assay, Mutagenesis, Plasmid Preparation, Pull Down Assay, Quantitative RT-PCR, Expressing, Transfection

    Silencing of circ‐CCND1 retards LSCC cell growth both in vitro and in vivo. A, qRT‐PCR analysis for circ‐CCND1 expression in LSCC cell lines. B, The diagram showing three designed siRNAs targeting the junction site of circ‐CCND1 (left). qRT‐PCR analysis verifying the knockdown efficiency of si‐circ‐CCND1#1 and #2 (right). C, The cell number counted at the indicated time after circ‐CCND1 depletion. D and E, The colony formation and cell cycle assays in circ‐CCND1‐depleted Hep‐2 and TU212 cells. F, The representative image showing the nude mice bearing tumours injected with cholesterol‐coupled control or si‐circ‐CCND1, the expression levels of circ‐CCND1 and the volume and weight of subcutaneous tumours in each group were analysed. * P
    Figure Legend Snippet: Silencing of circ‐CCND1 retards LSCC cell growth both in vitro and in vivo. A, qRT‐PCR analysis for circ‐CCND1 expression in LSCC cell lines. B, The diagram showing three designed siRNAs targeting the junction site of circ‐CCND1 (left). qRT‐PCR analysis verifying the knockdown efficiency of si‐circ‐CCND1#1 and #2 (right). C, The cell number counted at the indicated time after circ‐CCND1 depletion. D and E, The colony formation and cell cycle assays in circ‐CCND1‐depleted Hep‐2 and TU212 cells. F, The representative image showing the nude mice bearing tumours injected with cholesterol‐coupled control or si‐circ‐CCND1, the expression levels of circ‐CCND1 and the volume and weight of subcutaneous tumours in each group were analysed. * P

    Techniques Used: In Vitro, In Vivo, Quantitative RT-PCR, Expressing, Mouse Assay, Injection

    circ‐CCND1 is frequently overexpressed in LSCC and predicts poor prognosis. A, The flow chart showing the identification of circ‐CCND1 in LSCC. B‐D, qRT‐PCR analysis for the expression of circ‐CCND1, hsa_circ_0023304 and hsa_circ_0023305 in 25 paired LSCC and adjacent normal tissues (ANT). E, qRT‐PCR analysis for circ‐CCND1 expression in 101 pairs of LSCC and normal tissues. F, Kaplan‐Meier plotter showing the survival curve of LSCC patients with high and low circ‐CCND1 expression. *** P
    Figure Legend Snippet: circ‐CCND1 is frequently overexpressed in LSCC and predicts poor prognosis. A, The flow chart showing the identification of circ‐CCND1 in LSCC. B‐D, qRT‐PCR analysis for the expression of circ‐CCND1, hsa_circ_0023304 and hsa_circ_0023305 in 25 paired LSCC and adjacent normal tissues (ANT). E, qRT‐PCR analysis for circ‐CCND1 expression in 101 pairs of LSCC and normal tissues. F, Kaplan‐Meier plotter showing the survival curve of LSCC patients with high and low circ‐CCND1 expression. *** P

    Techniques Used: Quantitative RT-PCR, Expressing

    circ‐CCND1 regulates the stability of CCND1 mRNA. A, B, The mRNA expression and protein expression of CCND1 in control and circ‐CCND1‐silenced Hep‐2 and TU212 cells were detected by qRT‐PCR and Western blot assays, respectively. C, qRT‐PCR analysis for the different part of CCND1 mRNA in control and circ‐CCND1‐silenced Hep‐2 and TU212 cells. D, qRT‐PCR analysis for CCND1 mRNA expression in control and circ‐CCND1‐silenced Hep‐2 and TU212 cells at the indicated time after treatment with 1 mg/mL actinomycin D. E, qRT‐PCR analysis for CCND1 mRNA expression in 48 matched LSCC and normal tissues. F, The correlation between circ‐CCND1 and CCND1 mRNA expression in 48 LSCC tissues. G, The proliferative capabilities of circ‐CCND1‐silenced Hep‐2 and TU212 cells after transfection with CCND1 expression plasmid. ** P
    Figure Legend Snippet: circ‐CCND1 regulates the stability of CCND1 mRNA. A, B, The mRNA expression and protein expression of CCND1 in control and circ‐CCND1‐silenced Hep‐2 and TU212 cells were detected by qRT‐PCR and Western blot assays, respectively. C, qRT‐PCR analysis for the different part of CCND1 mRNA in control and circ‐CCND1‐silenced Hep‐2 and TU212 cells. D, qRT‐PCR analysis for CCND1 mRNA expression in control and circ‐CCND1‐silenced Hep‐2 and TU212 cells at the indicated time after treatment with 1 mg/mL actinomycin D. E, qRT‐PCR analysis for CCND1 mRNA expression in 48 matched LSCC and normal tissues. F, The correlation between circ‐CCND1 and CCND1 mRNA expression in 48 LSCC tissues. G, The proliferative capabilities of circ‐CCND1‐silenced Hep‐2 and TU212 cells after transfection with CCND1 expression plasmid. ** P

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Transfection, Plasmid Preparation

    65) Product Images from "MicroRNA-539 inhibits the progression of Wilms’ Tumor through downregulation of JAG1 and Notch1/3"

    Article Title: MicroRNA-539 inhibits the progression of Wilms’ Tumor through downregulation of JAG1 and Notch1/3

    Journal: Cancer Biomarkers

    doi: 10.3233/CBM-181972

    Overexpression of miR-539 suppressed the proliferation, migration and invasion of WT cells. (A) The miR-539 expression in SK-NEP-1 cell lines. (B) The miR-539 expressions were examined in SK-NEP-1 cells containing miR-539 mimics or inhibitor via qRT-PCR. (C, D) The cell proliferation was measured in cells containing miR-539 mimics or inhibitor via MTT assay. (E, F) Cell migration and invasion analysis in SK-NEP-1 cells containing miR-539 mimics or inhibitor was examined by Transwell assay. * P
    Figure Legend Snippet: Overexpression of miR-539 suppressed the proliferation, migration and invasion of WT cells. (A) The miR-539 expression in SK-NEP-1 cell lines. (B) The miR-539 expressions were examined in SK-NEP-1 cells containing miR-539 mimics or inhibitor via qRT-PCR. (C, D) The cell proliferation was measured in cells containing miR-539 mimics or inhibitor via MTT assay. (E, F) Cell migration and invasion analysis in SK-NEP-1 cells containing miR-539 mimics or inhibitor was examined by Transwell assay. * P

    Techniques Used: Over Expression, Migration, Expressing, Quantitative RT-PCR, MTT Assay, Transwell Assay

    Downregulation of miR-539 was identified in WT tissues.(A) The expressions of miR-539 in WT tissues detected via qRT-PCR. (B) Low miR-539 expression was correlated with shorter overall survival of WT patients. * P
    Figure Legend Snippet: Downregulation of miR-539 was identified in WT tissues.(A) The expressions of miR-539 in WT tissues detected via qRT-PCR. (B) Low miR-539 expression was correlated with shorter overall survival of WT patients. * P

    Techniques Used: Quantitative RT-PCR, Expressing

    66) Product Images from "miR-1254 inhibits cell proliferation, migration, and invasion by down-regulating Smurf1 in gastric cancer"

    Article Title: miR-1254 inhibits cell proliferation, migration, and invasion by down-regulating Smurf1 in gastric cancer

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-018-1262-x

    Smurf1 was up-regulated in GC tissues and cells and was confirmed to be a direct target gene of miR-1254. a Ninety pairs of GC tissues and adjacent normal tissues were collected to examine the relative expression of Smurf1 by qRT-PCR. b Western blot was used to determine Smurf1 protein expression in eight pairs GC specimens and adjacent normal tissues. c Immunohistochemistry staining was used to determine the protein level of Smurf1 in GC tissues and adjacent tissues (scale bar: 50 μm). d The correlation between the expression of miR-1254 and Smurf1. e and f The expression of Smurf1 in GC cells and GES-1 were detected by qRT-PCR and western blot, respectively. g Luciferase reporter assay was conducted to confirm that miR-1254 is directly bound to the 3′-UTR region of Smurf1. The result of Luciferase activity was analyzed in cells co-transfected with miR-1254-mimics or negative control with pGL3-Smurf1-WT or pGL3-Smurf1-MUT. h The expression level of Smurf1 protein in GC cells was verified by western blot. i The expression level of Smurf1 mRNA in GC cells was detected by qRT-PCR. * p
    Figure Legend Snippet: Smurf1 was up-regulated in GC tissues and cells and was confirmed to be a direct target gene of miR-1254. a Ninety pairs of GC tissues and adjacent normal tissues were collected to examine the relative expression of Smurf1 by qRT-PCR. b Western blot was used to determine Smurf1 protein expression in eight pairs GC specimens and adjacent normal tissues. c Immunohistochemistry staining was used to determine the protein level of Smurf1 in GC tissues and adjacent tissues (scale bar: 50 μm). d The correlation between the expression of miR-1254 and Smurf1. e and f The expression of Smurf1 in GC cells and GES-1 were detected by qRT-PCR and western blot, respectively. g Luciferase reporter assay was conducted to confirm that miR-1254 is directly bound to the 3′-UTR region of Smurf1. The result of Luciferase activity was analyzed in cells co-transfected with miR-1254-mimics or negative control with pGL3-Smurf1-WT or pGL3-Smurf1-MUT. h The expression level of Smurf1 protein in GC cells was verified by western blot. i The expression level of Smurf1 mRNA in GC cells was detected by qRT-PCR. * p

    Techniques Used: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry, Staining, Luciferase, Reporter Assay, Activity Assay, Transfection, Negative Control

    67) Product Images from "miR-564 inhibits hepatocellular carcinoma cell proliferation and invasion by targeting the GRB2-ERK1/2-AKT axis"

    Article Title: miR-564 inhibits hepatocellular carcinoma cell proliferation and invasion by targeting the GRB2-ERK1/2-AKT axis

    Journal: Oncotarget

    doi: 10.18632/oncotarget.22504

    GRB2 is upregulated in HCC and is inversely correlated with miR-564 expression ( A ) GRB2 mRNA expression in 20 HCC and adjacent noncancerous tissues as detected by qRT-PCR. ( B – C ) The correlation between miR-564 and GRB2 mRNA expression levels in cancer tissues and adjacent tissues was determined by Spearman's correlation analysis. ( D – F) . GRB2 was differentially expressed in the GSE14520 dataset, as determined using the Limma package on the R platform. The cutoff for significantly differentially expressed levels of miR-564 was P value
    Figure Legend Snippet: GRB2 is upregulated in HCC and is inversely correlated with miR-564 expression ( A ) GRB2 mRNA expression in 20 HCC and adjacent noncancerous tissues as detected by qRT-PCR. ( B – C ) The correlation between miR-564 and GRB2 mRNA expression levels in cancer tissues and adjacent tissues was determined by Spearman's correlation analysis. ( D – F) . GRB2 was differentially expressed in the GSE14520 dataset, as determined using the Limma package on the R platform. The cutoff for significantly differentially expressed levels of miR-564 was P value

    Techniques Used: Expressing, Quantitative RT-PCR

    miR-564 inhibits SMCC7721 and MHCC97H cell proliferation, migration and invasion in vitro ( A ) miR-564 expression in the miR-564 group and miR-NC group as measured by qRT-PCR. U6 was used as a loading control. There was a significant increase in the expression level of miR-564 in the miR-564 group ( P
    Figure Legend Snippet: miR-564 inhibits SMCC7721 and MHCC97H cell proliferation, migration and invasion in vitro ( A ) miR-564 expression in the miR-564 group and miR-NC group as measured by qRT-PCR. U6 was used as a loading control. There was a significant increase in the expression level of miR-564 in the miR-564 group ( P

    Techniques Used: Migration, In Vitro, Expressing, Quantitative RT-PCR

    GRB2 is a direct target of miR-564 in hepatocellular cells ( A ) The predicted miR-564 binding site in the 3′-UTR of GRB2 and the mutated 3′-UTR of GRB2, which was generated using the complementary sequence in the seed region of miR-564. ( B ) miR-564 or NC and a luciferase vector encoding the WT or MUT GRB2 3′-UTR were transfected into 293T cells, and the relative luciferase activity was measured. ( C ) GRB2 mRNA expression in the MHCC97H and SMCC7721 cells of the miR-564 and miR-NC groups as detected by qRT-PCR. ( D – E ) GRB2 protein expression in the MHCC97H and SMCC7721 cells of the miR-564 and miR-NC groups as detected by western blotting. All assays were repeated three times, and the mean values were used for comparisons.
    Figure Legend Snippet: GRB2 is a direct target of miR-564 in hepatocellular cells ( A ) The predicted miR-564 binding site in the 3′-UTR of GRB2 and the mutated 3′-UTR of GRB2, which was generated using the complementary sequence in the seed region of miR-564. ( B ) miR-564 or NC and a luciferase vector encoding the WT or MUT GRB2 3′-UTR were transfected into 293T cells, and the relative luciferase activity was measured. ( C ) GRB2 mRNA expression in the MHCC97H and SMCC7721 cells of the miR-564 and miR-NC groups as detected by qRT-PCR. ( D – E ) GRB2 protein expression in the MHCC97H and SMCC7721 cells of the miR-564 and miR-NC groups as detected by western blotting. All assays were repeated three times, and the mean values were used for comparisons.

    Techniques Used: Binding Assay, Generated, Sequencing, Luciferase, Plasmid Preparation, Transfection, Activity Assay, Expressing, Quantitative RT-PCR, Western Blot

    miR-564 expression is downregulated in HCC ( A ) miR-564 expression in HCC tissues and adjacent noncancerous tissues was quantified by qRT-PCR. The differences were statistically significant. ( B ) The expression of miR-564 in different HCC cell lines and the normal liver cell line LO-2 was measured by qRT-PCR; U6 was used as an internal reference. ( C ) miR-564 data were collected from the GEO dataset GSE21362. After quality control, 73 HCC and adjacent noncancerous liver samples were used for the analysis. ( D ) Differentially expressed levels of miR-564 in the GSE14520 dataset were determined using the Limma package on the R platform. The cutoff values for significantly differentially expressed miR-564 were a P value
    Figure Legend Snippet: miR-564 expression is downregulated in HCC ( A ) miR-564 expression in HCC tissues and adjacent noncancerous tissues was quantified by qRT-PCR. The differences were statistically significant. ( B ) The expression of miR-564 in different HCC cell lines and the normal liver cell line LO-2 was measured by qRT-PCR; U6 was used as an internal reference. ( C ) miR-564 data were collected from the GEO dataset GSE21362. After quality control, 73 HCC and adjacent noncancerous liver samples were used for the analysis. ( D ) Differentially expressed levels of miR-564 in the GSE14520 dataset were determined using the Limma package on the R platform. The cutoff values for significantly differentially expressed miR-564 were a P value

    Techniques Used: Expressing, Quantitative RT-PCR

    68) Product Images from "Hepatitis B virus X protein accelerates hepatocarcinogenesis with partner survivin through modulating miR-520b and HBXIP"

    Article Title: Hepatitis B virus X protein accelerates hepatocarcinogenesis with partner survivin through modulating miR-520b and HBXIP

    Journal: Molecular Cancer

    doi: 10.1186/1476-4598-13-128

    HBx down-regulates miR-520b through binding to Sp1 with partner survivin. (A) The expression levels of miRNA-520b, miRNA-520e, miRNA-29a and miRNA-181c were examined by qRT-PCR in LO2-X-S/LO2-X cells. (B) The expression of miR-520b in clinical HCC and peritumor samples was detected by qRT-PCR. (C) A model shows Sp1 binding site-directed mutation in the promoter region of miR-520b. (D) The effect of knockdown of Sp1 on miR-520b in LO2-X-S or HepG2.2.15 cells was examined by qRT-PCR analysis (*** P
    Figure Legend Snippet: HBx down-regulates miR-520b through binding to Sp1 with partner survivin. (A) The expression levels of miRNA-520b, miRNA-520e, miRNA-29a and miRNA-181c were examined by qRT-PCR in LO2-X-S/LO2-X cells. (B) The expression of miR-520b in clinical HCC and peritumor samples was detected by qRT-PCR. (C) A model shows Sp1 binding site-directed mutation in the promoter region of miR-520b. (D) The effect of knockdown of Sp1 on miR-520b in LO2-X-S or HepG2.2.15 cells was examined by qRT-PCR analysis (*** P

    Techniques Used: Binding Assay, Expressing, Quantitative RT-PCR, Mutagenesis

    MiR-520b/HBXIP modulates growth of LO2-X-S cells in vivo. (A-G) The effect of miR-520b or si-HBXIP on the growth of LO2-X-S cells was detected by xenograft assay. The expression levels of miR-520b and HBXIP in the tumor tissues from mice (n = 5, each group) were determined by qRT-PCR or western blot analysis, respectively. Data are shown as mean ± SD of three independent experiments (* P
    Figure Legend Snippet: MiR-520b/HBXIP modulates growth of LO2-X-S cells in vivo. (A-G) The effect of miR-520b or si-HBXIP on the growth of LO2-X-S cells was detected by xenograft assay. The expression levels of miR-520b and HBXIP in the tumor tissues from mice (n = 5, each group) were determined by qRT-PCR or western blot analysis, respectively. Data are shown as mean ± SD of three independent experiments (* P

    Techniques Used: In Vivo, Xenograft Assay, Expressing, Mouse Assay, Quantitative RT-PCR, Western Blot

    69) Product Images from "Tanshinone IIA pretreatment protects free flaps against hypoxic injury by upregulating stem cell-related biomarkers in epithelial skin cells"

    Article Title: Tanshinone IIA pretreatment protects free flaps against hypoxic injury by upregulating stem cell-related biomarkers in epithelial skin cells

    Journal: BMC Complementary and Alternative Medicine

    doi: 10.1186/1472-6882-14-331

    Epidermal cells showed upregulated Wnt signaling and enhanced stemness after pretreatment with TSA. qRT-PCR showed upregulation of β-catenin, SOX2, Nanog, OCT4, and VEGF and downregulation of GSK-3β in TSA-pretreated epithelial cells compared with control cells (A) . Similar results were obtained using western blot analysis (B) .
    Figure Legend Snippet: Epidermal cells showed upregulated Wnt signaling and enhanced stemness after pretreatment with TSA. qRT-PCR showed upregulation of β-catenin, SOX2, Nanog, OCT4, and VEGF and downregulation of GSK-3β in TSA-pretreated epithelial cells compared with control cells (A) . Similar results were obtained using western blot analysis (B) .

    Techniques Used: Quantitative RT-PCR, Western Blot

    70) Product Images from "RUNX2 Mutation Impairs 1α,25-Dihydroxyvitamin D3 mediated Osteoclastogenesis in Dental Follicle Cells"

    Article Title: RUNX2 Mutation Impairs 1α,25-Dihydroxyvitamin D3 mediated Osteoclastogenesis in Dental Follicle Cells

    Journal: Scientific Reports

    doi: 10.1038/srep24225

    Expressions of osteoclast-related genes were restrained by RUNX2 mutation in hDFCs and PBMCs co-culture and regulated by 1α,25-(OH) 2 D 3 stimulation. hDFCs from normal control ( RUNX2 +/+ ) and CCD patient ( RUNX2 +/m ) were co-cultured with peripheral blood mononuclear cells (PBMCs) in culture medium containing 1 × 10 −7 M 1α,25-(OH) 2 D 3 or equal volume of ethanol for 14 d (n = 4). ( a – e ) qRT-PCR was used to investigate the expression of RUNX2 and osteoclast-associated genes including CTR , TRAP, CTSK and MMP9 (n = 4). * P
    Figure Legend Snippet: Expressions of osteoclast-related genes were restrained by RUNX2 mutation in hDFCs and PBMCs co-culture and regulated by 1α,25-(OH) 2 D 3 stimulation. hDFCs from normal control ( RUNX2 +/+ ) and CCD patient ( RUNX2 +/m ) were co-cultured with peripheral blood mononuclear cells (PBMCs) in culture medium containing 1 × 10 −7 M 1α,25-(OH) 2 D 3 or equal volume of ethanol for 14 d (n = 4). ( a – e ) qRT-PCR was used to investigate the expression of RUNX2 and osteoclast-associated genes including CTR , TRAP, CTSK and MMP9 (n = 4). * P

    Techniques Used: Mutagenesis, Co-Culture Assay, Cell Culture, Quantitative RT-PCR, Expressing

    RUNX2 mutation affects the modulation effect of 1α,25-(OH) 2 D 3 on osteoclast-inductive capacities of hDFCs through RANKL/OPG signaling. hDFCs from normal control ( RUNX2 +/+ ) and CCD patient ( RUNX2 +/m ) were co-cultured with PBMCs in culture medium containing 1 × 10 −7 M 1α,25-(OH) 2 D 3 or equal volume of ethanol for 3 or 14 d (n = 4). ( a , b ) qRT-PCR was used to investigate the expression of RANKL and OPG after co-culturing for 14 d (n = 4). ( c ) The ratio of RANKL/OPG mRNA expression in hDFCs on 14 d (n = 4). ( d , e ) ELISA assay was used to analyze RANKL and OPG protein concentration in conditioned medium after co-culturing for 3 d (n = 4). ( f ) The ratio of RANKL/OPG protein concentration in conditioned medium on 14 d (n = 4). * P
    Figure Legend Snippet: RUNX2 mutation affects the modulation effect of 1α,25-(OH) 2 D 3 on osteoclast-inductive capacities of hDFCs through RANKL/OPG signaling. hDFCs from normal control ( RUNX2 +/+ ) and CCD patient ( RUNX2 +/m ) were co-cultured with PBMCs in culture medium containing 1 × 10 −7 M 1α,25-(OH) 2 D 3 or equal volume of ethanol for 3 or 14 d (n = 4). ( a , b ) qRT-PCR was used to investigate the expression of RANKL and OPG after co-culturing for 14 d (n = 4). ( c ) The ratio of RANKL/OPG mRNA expression in hDFCs on 14 d (n = 4). ( d , e ) ELISA assay was used to analyze RANKL and OPG protein concentration in conditioned medium after co-culturing for 3 d (n = 4). ( f ) The ratio of RANKL/OPG protein concentration in conditioned medium on 14 d (n = 4). * P

    Techniques Used: Mutagenesis, Cell Culture, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Protein Concentration

    71) Product Images from "MicroRNA Let-7g Directly Targets Forkhead Box C2 (FOXC2) to Modulate Bone Metastasis in Breast Cancer"

    Article Title: MicroRNA Let-7g Directly Targets Forkhead Box C2 (FOXC2) to Modulate Bone Metastasis in Breast Cancer

    Journal: Open Medicine

    doi: 10.1515/med-2017-0023

    Let-7g inhibits FOXC2 expression in breast cancer cells. (A) The levels of FOXC2 protein were examined in MDA-MB-231 cells with the treatment of let-7g by Western blot analysis. The successful transfection of let-7g was confirmed by qRT-PCR analysis. (B) The levels of FOXC2 protein were determined in SK-BR3 cells with anti-let-7g treatment using Western blot. The successful transfection of anti-let-7g was accessed by qRT-PCR. (Statistical differences: ** P
    Figure Legend Snippet: Let-7g inhibits FOXC2 expression in breast cancer cells. (A) The levels of FOXC2 protein were examined in MDA-MB-231 cells with the treatment of let-7g by Western blot analysis. The successful transfection of let-7g was confirmed by qRT-PCR analysis. (B) The levels of FOXC2 protein were determined in SK-BR3 cells with anti-let-7g treatment using Western blot. The successful transfection of anti-let-7g was accessed by qRT-PCR. (Statistical differences: ** P

    Techniques Used: Expressing, Multiple Displacement Amplification, Western Blot, Transfection, Quantitative RT-PCR

    Let-7g negatively correlates with FOXC2 in human breast cancer samples with bone metastasis. (A) FOXC2 expression was tested by qRT-PCR in 25 breast tumor tissues with bone metastasis and peritumor tissues. (B) The association of let-7g with FOXC2 was analyzed by qRT-PCR in 25 breast cancer tissues with bone metastasis (Pearson’s correlation coefficient, r=-0.8982). (Statistical differences: *** P
    Figure Legend Snippet: Let-7g negatively correlates with FOXC2 in human breast cancer samples with bone metastasis. (A) FOXC2 expression was tested by qRT-PCR in 25 breast tumor tissues with bone metastasis and peritumor tissues. (B) The association of let-7g with FOXC2 was analyzed by qRT-PCR in 25 breast cancer tissues with bone metastasis (Pearson’s correlation coefficient, r=-0.8982). (Statistical differences: *** P

    Techniques Used: Expressing, Quantitative RT-PCR

    72) Product Images from "LncRNA NEAT1 contributes to paclitaxel resistance of ovarian cancer cells by regulating ZEB1 expression via miR-194"

    Article Title: LncRNA NEAT1 contributes to paclitaxel resistance of ovarian cancer cells by regulating ZEB1 expression via miR-194

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S147586

    NEAT1 knockdown-sensitized SKOV3/PTX and HeyA-8/PTX cells to PTX. Notes: IC 50 value of PTX in SKOV3 and SKOV3/PTX cells ( A ) as well as in HeyA-8 and HeyA-8/PTX cells ( B ). Expression of NEAT1 in si-NEAT1- or si-con-transfected SKOV3/PTX ( C ) and HeyA-8/PTX ( D ) cells was estimated by qRT-PCR. Protein levels of P-gp and GST-π were determined by Western blot in SKOV3/PTX ( E ) and HeyA-8/PTX ( F ) cells after introduction with si-NEAT1 or si-con. PTX resistance of si-NEAT1- or si-con-treated SKOV3/PTX ( G ) and HeyA-8/PTX ( H ) cells was assessed using IC 50 value of PTX by MTT assay. Apoptotic rate was measured by flow cytometry after SKOV3/PTX ( I ) and HeyA-8/PTX ( J ) cells transfected with si-NEAT1 or si-con were treated with PTX for 48 h. * P
    Figure Legend Snippet: NEAT1 knockdown-sensitized SKOV3/PTX and HeyA-8/PTX cells to PTX. Notes: IC 50 value of PTX in SKOV3 and SKOV3/PTX cells ( A ) as well as in HeyA-8 and HeyA-8/PTX cells ( B ). Expression of NEAT1 in si-NEAT1- or si-con-transfected SKOV3/PTX ( C ) and HeyA-8/PTX ( D ) cells was estimated by qRT-PCR. Protein levels of P-gp and GST-π were determined by Western blot in SKOV3/PTX ( E ) and HeyA-8/PTX ( F ) cells after introduction with si-NEAT1 or si-con. PTX resistance of si-NEAT1- or si-con-treated SKOV3/PTX ( G ) and HeyA-8/PTX ( H ) cells was assessed using IC 50 value of PTX by MTT assay. Apoptotic rate was measured by flow cytometry after SKOV3/PTX ( I ) and HeyA-8/PTX ( J ) cells transfected with si-NEAT1 or si-con were treated with PTX for 48 h. * P

    Techniques Used: Expressing, Transfection, Quantitative RT-PCR, Western Blot, MTT Assay, Flow Cytometry, Cytometry

    NEAT1 inhibited miR-194 expression. Notes: ( A ) Graphical representation of predicted wild-type or mutated binding sequences in NEAT1. Luciferase reporter assay in SKOV3/PTX ( B ) and HeyA-8/PTX ( C ) cells after cotransfection with established luciferase reporter vectors (pGL3-NEAT1-Wt or pGL3-NEAT1-Mut) and miR-194 or miR-con. Expression of miR-194 was examined by qRT-PCR in SKOV3/PTX ( D ) and HeyA-8/PTX ( E ) cells treated with pcDNA-NEAT1, si-NEAT1, or respective controls. * P
    Figure Legend Snippet: NEAT1 inhibited miR-194 expression. Notes: ( A ) Graphical representation of predicted wild-type or mutated binding sequences in NEAT1. Luciferase reporter assay in SKOV3/PTX ( B ) and HeyA-8/PTX ( C ) cells after cotransfection with established luciferase reporter vectors (pGL3-NEAT1-Wt or pGL3-NEAT1-Mut) and miR-194 or miR-con. Expression of miR-194 was examined by qRT-PCR in SKOV3/PTX ( D ) and HeyA-8/PTX ( E ) cells treated with pcDNA-NEAT1, si-NEAT1, or respective controls. * P

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

    Expressions of NEAT1 and miR-194 in PTX-resistant ovarian cancer tissues and cells. Notes: qRT-PCR analysis of NEAT1 ( A ) and miR-194 ( B ) expressions in 18 treatment-responsive patients and 14 treatment-resistant patients. qRT-PCR analysis of NEAT1 ( C and D ) and miR-194 ( E and F ) expressions in parental ovarian cancer cells (SKOV3 and HeyA-8) and PTX-resistant ovarian cancer cells (SKOV3/PTX and HeyA-8/PTX). * P
    Figure Legend Snippet: Expressions of NEAT1 and miR-194 in PTX-resistant ovarian cancer tissues and cells. Notes: qRT-PCR analysis of NEAT1 ( A ) and miR-194 ( B ) expressions in 18 treatment-responsive patients and 14 treatment-resistant patients. qRT-PCR analysis of NEAT1 ( C and D ) and miR-194 ( E and F ) expressions in parental ovarian cancer cells (SKOV3 and HeyA-8) and PTX-resistant ovarian cancer cells (SKOV3/PTX and HeyA-8/PTX). * P

    Techniques Used: Quantitative RT-PCR

    73) Product Images from "Downregulation of microRNA-195 promotes angiogenesis induced by cerebral infarction via targeting VEGFA"

    Article Title: Downregulation of microRNA-195 promotes angiogenesis induced by cerebral infarction via targeting VEGFA

    Journal: Molecular Medicine Reports

    doi: 10.3892/mmr.2017.7230

    miR-195 regulates migration and tube formation of HUVECs. (A) HUVECs were treated with hypoxia for the indicated time-points. The relative expression level of miR-195 was detected by qPT-PCR. (B) miR-195 mimics and miR-195 inhibitor were transfected into HUVECs. After 24 h, cells were treated with hypoxia for 12 h. The relative expression level of miR-195 was detected by qRT-PCR. (C and E) HUVECs were transfected with miR-195 mimic, miR-195 inhibitor or miR mimic scramble. After 24 h, cells were treated with hypoxia for 12 h. The tube formation assay was performed as described in Materials and methods. The length of the tubes was quantified. (D and F) HUVECs were transfected as in C and E, and then cells were treated with hypoxia for 24 h. The cell migration assay was performed a s described in Materials and methods. The histogram represents the quantification of cells that migrated. All data are expressed as the mean ± SD. *P
    Figure Legend Snippet: miR-195 regulates migration and tube formation of HUVECs. (A) HUVECs were treated with hypoxia for the indicated time-points. The relative expression level of miR-195 was detected by qPT-PCR. (B) miR-195 mimics and miR-195 inhibitor were transfected into HUVECs. After 24 h, cells were treated with hypoxia for 12 h. The relative expression level of miR-195 was detected by qRT-PCR. (C and E) HUVECs were transfected with miR-195 mimic, miR-195 inhibitor or miR mimic scramble. After 24 h, cells were treated with hypoxia for 12 h. The tube formation assay was performed as described in Materials and methods. The length of the tubes was quantified. (D and F) HUVECs were transfected as in C and E, and then cells were treated with hypoxia for 24 h. The cell migration assay was performed a s described in Materials and methods. The histogram represents the quantification of cells that migrated. All data are expressed as the mean ± SD. *P

    Techniques Used: Migration, Expressing, Polymerase Chain Reaction, Transfection, Quantitative RT-PCR, Tube Formation Assay, Cell Migration Assay

    74) Product Images from "Aldosterone signaling regulates the over-expression of claudin-4 and -8 at the distal nephron from type 1 diabetic rats"

    Article Title: Aldosterone signaling regulates the over-expression of claudin-4 and -8 at the distal nephron from type 1 diabetic rats

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0177362

    SPL treatment blunts diabetes-induced increment of protein and mRNA expressions of cldn-4 and -8. SPL decreased diabetes-induced cldn-4 and -8 expressions evaluated by IF, IB and qRT- PCR. IF analysis shows that 3 weeks after diabetes induction, increased expressions of (A) cldn-4 (green label) and (E) cldn-8 (green label) were found in the TJs of DT labeled with desmoplakin (DMPK, red label). Nuclei were marked with DAPI (blue label). SPL treatment significantly decreased these changes. To confirm IF findings, IB and qRT-PCR analyzes were performed ( S4 Fig ). Diabetic condition significantly increased protein (B) and mRNA (D) levels of cldn-4; these changes were decreased by SPL treatment. Also, SPL treatment decreased diabetes-induced increased protein (F) and mRNA (H) levels of cldn-8. Similar results were found between CTL and SPL groups. GAPDH was used as loading control. Densitometric analyzes of IB from the four experimental groups are shown in panels C for cldn-4 and, G for cldn-8 ( S4 Fig ). Data are mean±SEM from 3 rats per group. *p
    Figure Legend Snippet: SPL treatment blunts diabetes-induced increment of protein and mRNA expressions of cldn-4 and -8. SPL decreased diabetes-induced cldn-4 and -8 expressions evaluated by IF, IB and qRT- PCR. IF analysis shows that 3 weeks after diabetes induction, increased expressions of (A) cldn-4 (green label) and (E) cldn-8 (green label) were found in the TJs of DT labeled with desmoplakin (DMPK, red label). Nuclei were marked with DAPI (blue label). SPL treatment significantly decreased these changes. To confirm IF findings, IB and qRT-PCR analyzes were performed ( S4 Fig ). Diabetic condition significantly increased protein (B) and mRNA (D) levels of cldn-4; these changes were decreased by SPL treatment. Also, SPL treatment decreased diabetes-induced increased protein (F) and mRNA (H) levels of cldn-8. Similar results were found between CTL and SPL groups. GAPDH was used as loading control. Densitometric analyzes of IB from the four experimental groups are shown in panels C for cldn-4 and, G for cldn-8 ( S4 Fig ). Data are mean±SEM from 3 rats per group. *p

    Techniques Used: Quantitative RT-PCR, Labeling, CTL Assay

    SPL treatment decreases diabetes-induced increased WNK4 expression and co-IP with cldn- 4 and -8. WNK4 expression was evaluated by IB and qRT-PCR in the four experimental groups ( S6 Fig ). As shown in panels A and C, diabetes induced WNK4 protein expression and mRNA levels, respectively. SPL treatment decreased these changes. Densitometric analysis of IB is shown in panel B. To evaluate direct interaction of WNK4 with cldn-4 and -8 at the TJ, co-IP analyzes were performed. As shown, diabetes induced co-IP of WNK4 with cldn-4 (D) and vice versa (E). Also, diabetic condition induced co-IP of WNK4 with cldn-8 (F) and vice versa (G). SPL treatment decreased these changes. As shown in panels D-G, no signal was found under nonspecific conditions of IP performed with an unrelated antibody. GAPDH was evaluated in isolated DT and input extract as loading control. Data are mean±SEM from 3 rats per group. *p
    Figure Legend Snippet: SPL treatment decreases diabetes-induced increased WNK4 expression and co-IP with cldn- 4 and -8. WNK4 expression was evaluated by IB and qRT-PCR in the four experimental groups ( S6 Fig ). As shown in panels A and C, diabetes induced WNK4 protein expression and mRNA levels, respectively. SPL treatment decreased these changes. Densitometric analysis of IB is shown in panel B. To evaluate direct interaction of WNK4 with cldn-4 and -8 at the TJ, co-IP analyzes were performed. As shown, diabetes induced co-IP of WNK4 with cldn-4 (D) and vice versa (E). Also, diabetic condition induced co-IP of WNK4 with cldn-8 (F) and vice versa (G). SPL treatment decreased these changes. As shown in panels D-G, no signal was found under nonspecific conditions of IP performed with an unrelated antibody. GAPDH was evaluated in isolated DT and input extract as loading control. Data are mean±SEM from 3 rats per group. *p

    Techniques Used: Expressing, Co-Immunoprecipitation Assay, Quantitative RT-PCR, Isolation

    SPL treatment decreases diabetes-induced decrement in protein expression of cldn-5 in glomeruli (GL) and cldn-2 and occldn in proximal tubules (PT). SPL prevents diabetes-induced decrease of cldn-5 protein expression (A and B) in GL, and mRNA (F) and protein expression of cldn-2 (A and C), and protein expression of occldn (A and D) in PT ( S2 Fig ). However, diabetes did not have effect in the mRNA levels of cldn-5 (E) and occldn (G) evaluated by qRT-PCR ( S2 Fig ). Additionally, cellular localization of cldn-2 and -5 and occldn were assessed by IF (H). SPL-treatment prevents diabetes-induced loss of cldn-5 (green label) in the cell borders of GL and, cldn-2 (green label) and occldn (green label) in the cell borders of PT labeled with DppD (red label), nuclei were marked with DAPI (blue label). No changes were found between CTL and SPL groups. GAPDH was used as loading control. Densitometric analysis of IBs from the four experimental groups are shown in panels B for cldn-5, C for cldn-2 and D for occldn. Data are mean±SEM from 3 rats per group. *p
    Figure Legend Snippet: SPL treatment decreases diabetes-induced decrement in protein expression of cldn-5 in glomeruli (GL) and cldn-2 and occldn in proximal tubules (PT). SPL prevents diabetes-induced decrease of cldn-5 protein expression (A and B) in GL, and mRNA (F) and protein expression of cldn-2 (A and C), and protein expression of occldn (A and D) in PT ( S2 Fig ). However, diabetes did not have effect in the mRNA levels of cldn-5 (E) and occldn (G) evaluated by qRT-PCR ( S2 Fig ). Additionally, cellular localization of cldn-2 and -5 and occldn were assessed by IF (H). SPL-treatment prevents diabetes-induced loss of cldn-5 (green label) in the cell borders of GL and, cldn-2 (green label) and occldn (green label) in the cell borders of PT labeled with DppD (red label), nuclei were marked with DAPI (blue label). No changes were found between CTL and SPL groups. GAPDH was used as loading control. Densitometric analysis of IBs from the four experimental groups are shown in panels B for cldn-5, C for cldn-2 and D for occldn. Data are mean±SEM from 3 rats per group. *p

    Techniques Used: Expressing, Quantitative RT-PCR, Labeling, CTL Assay

    75) Product Images from "Transcription factors regulate GPR91-mediated expression of VEGF in hypoxia-induced retinopathy"

    Article Title: Transcription factors regulate GPR91-mediated expression of VEGF in hypoxia-induced retinopathy

    Journal: Scientific Reports

    doi: 10.1038/srep45807

    Identification of functional binding sites for C/EBP and AP-1. ( A ) The potential binding sites for C/EBP and AP-1 transcription factors in the 2000-bp promoter sequence upstream of the VEGF gene. The numbers indicate the base location relative to the transcription start site. TSS: transcription start site. ( B ) Luciferase reports showing changes in VEGF promoter activity after mutations of the binding region using HEK293 incubated with 400 μM CoCl 2 for 24 h. ( C ) A ChIP assay (using qRT-PCR) for C/EBP β and C/EBP δ using retinal ganglion cells incubated with 200 μM CoCl 2 for 24 h. ( D ) A ChIP assay (using qRT-PCR) for c-Fos using retinal ganglion cells incubated with 200 μM CoCl 2 for 24 h. Each column denotes the mean ± SD (n = 3). ** P
    Figure Legend Snippet: Identification of functional binding sites for C/EBP and AP-1. ( A ) The potential binding sites for C/EBP and AP-1 transcription factors in the 2000-bp promoter sequence upstream of the VEGF gene. The numbers indicate the base location relative to the transcription start site. TSS: transcription start site. ( B ) Luciferase reports showing changes in VEGF promoter activity after mutations of the binding region using HEK293 incubated with 400 μM CoCl 2 for 24 h. ( C ) A ChIP assay (using qRT-PCR) for C/EBP β and C/EBP δ using retinal ganglion cells incubated with 200 μM CoCl 2 for 24 h. ( D ) A ChIP assay (using qRT-PCR) for c-Fos using retinal ganglion cells incubated with 200 μM CoCl 2 for 24 h. Each column denotes the mean ± SD (n = 3). ** P

    Techniques Used: Functional Assay, Binding Assay, Sequencing, Luciferase, Activity Assay, Incubation, Chromatin Immunoprecipitation, Quantitative RT-PCR

    GPR91 modulated the CoCl 2 -induced increase in ERK1/2 signaling, C/EBP β, c-Fos, HIF-1α activity and VEGF expression in retinal ganglion cells. ( A ) Changes in ERK1/2 phosphorylation (using western blot) in retinal ganglion cells transduced with LV.shScrambled or LV. shGPR91. ( B ) Quantitative analysis of band density. Each column denotes the mean ± SD (n = 3). ( C ) qRT-PCR analysis for C/EBP β and c-Fos mRNA in retinal ganglion cells from each group. ( D ) Western blot analysis of C/EBP β, c-Fos and HIF-1α protein expression in retinal ganglion cells treated with CoCl 2 for 24 h. The cells were transduced with GPR91 siRNA or pre-treated with U0126 (ERK1/2 inhibitor). ( E ) Quantitative analysis of band density. ( F ) Changes in mRNA levels of HIF-1αVEGF in samples from each group. ( G ) ELISA analysis of VEGF secretion in samples from each group. ( H ) Western blot analysis of VEGF secretion in samples from each group. ( I ) Quantitative analysis of band density. Each column denotes the mean ± SD (n = 3). Each column denotes the mean ± SD (n = 3). ** P
    Figure Legend Snippet: GPR91 modulated the CoCl 2 -induced increase in ERK1/2 signaling, C/EBP β, c-Fos, HIF-1α activity and VEGF expression in retinal ganglion cells. ( A ) Changes in ERK1/2 phosphorylation (using western blot) in retinal ganglion cells transduced with LV.shScrambled or LV. shGPR91. ( B ) Quantitative analysis of band density. Each column denotes the mean ± SD (n = 3). ( C ) qRT-PCR analysis for C/EBP β and c-Fos mRNA in retinal ganglion cells from each group. ( D ) Western blot analysis of C/EBP β, c-Fos and HIF-1α protein expression in retinal ganglion cells treated with CoCl 2 for 24 h. The cells were transduced with GPR91 siRNA or pre-treated with U0126 (ERK1/2 inhibitor). ( E ) Quantitative analysis of band density. ( F ) Changes in mRNA levels of HIF-1αVEGF in samples from each group. ( G ) ELISA analysis of VEGF secretion in samples from each group. ( H ) Western blot analysis of VEGF secretion in samples from each group. ( I ) Quantitative analysis of band density. Each column denotes the mean ± SD (n = 3). Each column denotes the mean ± SD (n = 3). ** P

    Techniques Used: Activity Assay, Expressing, Western Blot, Transduction, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    Distribution of GPR91 in the retinal ganglion cells. ( A ) Western blot analysis of GPR91 protein expression in retinal ganglion cells treated with CoCl 2 for 24 h. ( B ) Quantitative analysis of band density. Each column denotes the mean ± SD (n = 3). ( C ) Immunofluorescence showing GPR91 expression in the cytoplasm of retinal ganglion cells. Green fluorescence shows the distribution of the GPR91 protein. Red fluorescence shows the distribution of the retinal ganglion cell marker Thy1.1 protein. Blue fluorescence shows nuclei stained with 4,6-diamidino-2-phenylindole (DAPI). The right column shows the merged pictures. ( D ) Changes in mRNA levels (as determined using qRT-PCR) of GPR91 in samples from each group. ( E ) Western blot analysis of the GPR91 protein in samples from each group. ( F ) Quantitative analysis of the band density of GPR91/β-actin. Each column denotes the mean ± SD (n = 3). ** P
    Figure Legend Snippet: Distribution of GPR91 in the retinal ganglion cells. ( A ) Western blot analysis of GPR91 protein expression in retinal ganglion cells treated with CoCl 2 for 24 h. ( B ) Quantitative analysis of band density. Each column denotes the mean ± SD (n = 3). ( C ) Immunofluorescence showing GPR91 expression in the cytoplasm of retinal ganglion cells. Green fluorescence shows the distribution of the GPR91 protein. Red fluorescence shows the distribution of the retinal ganglion cell marker Thy1.1 protein. Blue fluorescence shows nuclei stained with 4,6-diamidino-2-phenylindole (DAPI). The right column shows the merged pictures. ( D ) Changes in mRNA levels (as determined using qRT-PCR) of GPR91 in samples from each group. ( E ) Western blot analysis of the GPR91 protein in samples from each group. ( F ) Quantitative analysis of the band density of GPR91/β-actin. Each column denotes the mean ± SD (n = 3). ** P

    Techniques Used: Western Blot, Expressing, Immunofluorescence, Fluorescence, Marker, Staining, Quantitative RT-PCR

    CoCl 2 -induced activation of ERK1/2 signaling, C/EBP and AP-1 in retinal ganglion cells. ( A ) Western blot analysis of ERK1/2 phosphorylation in retinal ganglion cells treated with 200 μM CoCl 2 for different periods of time. ( B ) Quantitative analysis of the band density of p-ERK1/2/t-ERK1/2. ( C ) qRT-PCR analysis of C/EBP subtypes in retinal ganglion cells treated with 200 μM CoCl 2 for 24 h. ( D ) qRT-PCR analysis of AP-1 subtypes in retinal ganglion cells treated with 200 μM CoCl 2 for 24 h. ( E ) Changes in C/EBP β, C/EBP δ and c-Fos protein levels in CoCl 2 -induced retinal ganglion cells treated for 24 h. ( F – H ) Quantitative analysis of the band density. Each column denotes the mean ± SD (n = 3). ** P
    Figure Legend Snippet: CoCl 2 -induced activation of ERK1/2 signaling, C/EBP and AP-1 in retinal ganglion cells. ( A ) Western blot analysis of ERK1/2 phosphorylation in retinal ganglion cells treated with 200 μM CoCl 2 for different periods of time. ( B ) Quantitative analysis of the band density of p-ERK1/2/t-ERK1/2. ( C ) qRT-PCR analysis of C/EBP subtypes in retinal ganglion cells treated with 200 μM CoCl 2 for 24 h. ( D ) qRT-PCR analysis of AP-1 subtypes in retinal ganglion cells treated with 200 μM CoCl 2 for 24 h. ( E ) Changes in C/EBP β, C/EBP δ and c-Fos protein levels in CoCl 2 -induced retinal ganglion cells treated for 24 h. ( F – H ) Quantitative analysis of the band density. Each column denotes the mean ± SD (n = 3). ** P

    Techniques Used: Activation Assay, Western Blot, Quantitative RT-PCR

    Inhibition of VEGF expression in the retinas of OIR rats following intravitreal injection of LV.shGPR91 particles. ( A ) qRT-PCR analysis of VEGF mRNA in P18 OIR rat retinas and age-matched RA rats. ( B ) qRT-PCR analysis for GPR91 mRNA in P18 OIR rat retinas and age-matched RA rats. ( C ) Western blot analysis of the GPR91 and VEGF proteins in samples from each group. ( D , E ) Quantitative analysis of banddensities. Each column denotes the mean ± SD (n = 6). ** P
    Figure Legend Snippet: Inhibition of VEGF expression in the retinas of OIR rats following intravitreal injection of LV.shGPR91 particles. ( A ) qRT-PCR analysis of VEGF mRNA in P18 OIR rat retinas and age-matched RA rats. ( B ) qRT-PCR analysis for GPR91 mRNA in P18 OIR rat retinas and age-matched RA rats. ( C ) Western blot analysis of the GPR91 and VEGF proteins in samples from each group. ( D , E ) Quantitative analysis of banddensities. Each column denotes the mean ± SD (n = 6). ** P

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

    GPR91 modulated the increase in VEGF via the ERK1/2/ C/EBP β (c-Fos, HIF-1α) pathway in OIR rats. ( A ) Western blot analysis of ERK1/2 signaling, C/EBP β, c-Fos and HIF-1α activation in samples from each group. ( B ) Quantitative analysis of band density. ( C ) qRT-PCR analysis for VEGF mRNA in retinal ganglion cells from each group. ( D ) Western blot analysis of VEGF expression in samples from each group. ( E ) Quantitative analysis of the band density. Each column denotes the mean ± SD (n = 3). ** P
    Figure Legend Snippet: GPR91 modulated the increase in VEGF via the ERK1/2/ C/EBP β (c-Fos, HIF-1α) pathway in OIR rats. ( A ) Western blot analysis of ERK1/2 signaling, C/EBP β, c-Fos and HIF-1α activation in samples from each group. ( B ) Quantitative analysis of band density. ( C ) qRT-PCR analysis for VEGF mRNA in retinal ganglion cells from each group. ( D ) Western blot analysis of VEGF expression in samples from each group. ( E ) Quantitative analysis of the band density. Each column denotes the mean ± SD (n = 3). ** P

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

    Related Articles

    Countercurrent Chromatography:

    Article Title: Antinociceptive Effect of Tetrandrine on LPS-Induced Hyperalgesia via the Inhibition of IKK? Phosphorylation and the COX-2/PGE2 Pathway in Mice
    Article Snippet: 2.9 Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was isolated from brain tissues and cultured cells using Trizol reagent, (Invitrogen, USA) according to the manufacturer's guidelines, followed by further purification using the RNeasy Mini Kits (Qiagen, USA). .. The following oligonucleotide primers were used: COX-1 forward: 5′-TGC CCT CTG TAC CCA AAG AC-3′ , reverse: 5′-GGA CCC ATC TTT CCA GAG GT-3′ ; COX-2 forward: 5′-CGG AGA GAG TTC ATC CCT GA-3′ ; reverse: 5′-ATC CTT GAA AAG GCG CAG T-3′ ; GAPDH (control) forward: 5′-CTC ATG ACC ACA GTC CAT GC-3′ , reverse: 5′-CAC ATT GGG GGT AGG AAC AC-3′ .

    Real-time Polymerase Chain Reaction:

    Article Title: Antinociceptive Effect of Tetrandrine on LPS-Induced Hyperalgesia via the Inhibition of IKK? Phosphorylation and the COX-2/PGE2 Pathway in Mice
    Article Snippet: .. 2.9 Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was isolated from brain tissues and cultured cells using Trizol reagent, (Invitrogen, USA) according to the manufacturer's guidelines, followed by further purification using the RNeasy Mini Kits (Qiagen, USA). .. Purified RNA was reverse-transcribed into cDNA with random hexamers by the SuperScriptTM First-Strand Synthesis System kit (Qiagen, USA) and analyzed by real-time RT-PCR with the QuantiTect SYBR Green PCR Kits (Qiagen, USA), using a MJ DNA Engine Opticon2 qPCR System (MJ, USA).

    Article Title: Targeting Gastrin-Releasing Peptide Suppresses Neuroblastoma Progression via Upregulation of PTEN Signaling
    Article Snippet: .. Reverse transcription, semi-quantitative and quantitative real-time polymerase chain reaction (QRT-PCR) Total RNA was isolated using Trizol and reverse-transcribed to cDNA using High Capacity cDNA reverse transcription kit according to manufacturer's protocol (Applied Biosystems, Foster City, CA). .. Semi-quantitative PCR was performed using a Peltier Thermal Cycler (PTC-200) using specific 3′and 5′ primers for GRP and final product was visualized on 1% agarose gel using a Gel Doc (Bio-Rad).

    Article Title: Analysis of the miRNA Expression Profiles in the Zearalenone-Exposed TM3 Leydig Cell Line
    Article Snippet: .. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was extracted using Trizol reagent (Invitrogen, Carlsbad, CA, USA) following the manufacturer’s procedure. .. The total RNAs from each sample were reverse-transcribed to cDNA using the miRNA first-strand cDNA synthesis kit (by stem-loop) (Vazyme, Nanjing, China).

    Article Title: HSP90 inhibition downregulates thymidylate synthase and sensitizes colorectal cancer cell lines to the effect of 5FU-based chemotherapy
    Article Snippet: .. Quantitative real time-polymerase chain reaction (qRT-PCR) Total RNA was isolated using TRIzol reagent (Invitrogen Corporation, Carlsbad, CA). .. The reverse transcription (RT) step was performed with MultiScribeTM reverse transcriptase (Applied Biosystems, USA).

    Article Title: Resveratrol Exerts Dosage-Dependent Effects on the Self-Renewal and Neural Differentiation of hUC-MSCs
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was isolated using Trizol reagent (Invitrogen) according to the instructions, and mRNA was reverse transcribed using PrimeScript™ RT-PCR Kit (TaKaRa, Japan). qRT-PCR were performed on an ABI 7500 real-time PCR system (Thermo Fisher Scientific, USA) using a SYBR Premix Ex Taq™ (Prefect Real Time, Takara, Japan). ..

    Article Title: Annexin A2 Coordinates STAT3 to Regulate the Invasion and Migration of Colorectal Cancer Cells In Vitro
    Article Snippet: .. RNA Extraction and Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA of tissue samples was extracted by using the RecoverAll™ Total Nucleic Acid Isolation Kit (Ambion, Austin, USA). .. RNeasy kits (Qiagen, Valencia, CA) were utilized to extract total mRNA from cells. qRT-PCR was performed on ABI 7500 with SYBR Premix Ex Taq™ Kit (Takara, Japan).

    Article Title: Protein phosphatase 2A-B55δ enhances chemotherapy sensitivity of human hepatocellular carcinoma under the regulation of microRNA-133b
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted using TRIzol® Reagent (Ambion, TX, USA) and reverse-transcribed into cDNA using PrimeScript™ RT reagent kit (TaKaRa, Otsu, Japan). qRT-PCR was performed with SYBR® Premix Ex Taq ™ II kit (TaKaRa) in a CFX96 Touch™ Real-Time PCR Detection System (Bio-Rad, CA, USA) using the primers (Sangon Biotech, Shanghai, China) shown in Additional file : Table S1. ..

    Article Title: Exosomal miR-21-5p derived from gastric cancer promotes peritoneal metastasis via mesothelial-to-mesenchymal transition
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted from tissues, cultured cells and exosomes using the TRIzol reagent (Invitrogen) according to the manufacturer’s instructions. .. Then RNA was reverse transcribed into cDNA using PrimeScript RT Reagent (TaKaRa, Japan).

    Article Title: Strigolactone analogues induce apoptosis through activation of p38 and the stress response pathway in cancer cell lines and in conditionally reprogramed primary prostate cancer cells.
    Article Snippet: .. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was extracted using Trizol (Invitrogen, Carlsbad, CA, USA) using the manufacturer’s protocol. .. 1μg RNA was reverse-transcribed in a total volume of 20 μl using the High Capacity cDNA kit (Invitrogen).

    Article Title: miR-96 exerts carcinogenic effect by activating AKT/GSK-3β/β-catenin signaling pathway through targeting inhibition of FOXO1 in hepatocellular carcinoma
    Article Snippet: .. Quantitative real time polymerase chain reaction (qRT-PCR) Total RNA in tumor tissues and cells was obtained by Trizol kit (Thermo Fisher Scientific, Waltham, MA, USA) according to the the instruction manual. ..

    Article Title: Overexpression of Cullin7 is associated with hepatocellular carcinoma progression and pathogenesis
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted using TRIzol reagent (Invitrogen). .. As previously described [ ], First-strand cDNA was synthesized with a Super ScriptIII First-Strand Synthesis System (Invitrogen). qRT-PCR for cullin7 mRNA was performed on Applied Biosystems 7500 Fast Real-Time PCR system (Applied Biosystems) using One Step SYBR PrimeScript Plus RT-PCR kit (Takara Bio, Inc., Shiga, Japan) according to the manufacturer’s instruction.

    Article Title: LINC00460 modulates KDM2A to promote cell proliferation and migration by targeting miR-342-3p in gastric cancer
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted from tissues and cells using the Trizol reagent (Invitrogen) according to the manufacturer’s instructions. .. For microRNA analysis, qRT-PCR was performed using the TaqMan MicroRNA Reverse Transcription Kit, TaqMan Universal PCR Master Mix (Thermo Fisher Scientific), and the corresponding primers.

    Article Title: Doxycycline Attenuates Leptospira-Induced IL-1β by Suppressing NLRP3 Inflammasome Priming
    Article Snippet: .. Total RNA Isolation and Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA of cells and organs were extracted using TRIzol (Invitrogen, USA) following the manufacturer’s instructions. .. RNA was reverse transcribed into cDNA by using random primers from a TransScript One-Step gDNA Removal kit and cDNA Synthesis SuperMix (TransGen Biotech, China).

    Article Title: miR-22 inhibits mouse ovarian granulosa cell apoptosis by targeting SIRT1
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA were extracted from ovarian GCs of mice using a TRIZOL regent kit (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions. .. The purity of RNA was assessed by a NanoDrop 2000 UV-Vis spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA) at 260/280.

    RNA Extraction:

    Article Title: Annexin A2 Coordinates STAT3 to Regulate the Invasion and Migration of Colorectal Cancer Cells In Vitro
    Article Snippet: .. RNA Extraction and Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA of tissue samples was extracted by using the RecoverAll™ Total Nucleic Acid Isolation Kit (Ambion, Austin, USA). .. RNeasy kits (Qiagen, Valencia, CA) were utilized to extract total mRNA from cells. qRT-PCR was performed on ABI 7500 with SYBR Premix Ex Taq™ Kit (Takara, Japan).

    Agarose Gel Electrophoresis:

    Article Title: Targeting Gastrin-Releasing Peptide Suppresses Neuroblastoma Progression via Upregulation of PTEN Signaling
    Article Snippet: Reverse transcription, semi-quantitative and quantitative real-time polymerase chain reaction (QRT-PCR) Total RNA was isolated using Trizol and reverse-transcribed to cDNA using High Capacity cDNA reverse transcription kit according to manufacturer's protocol (Applied Biosystems, Foster City, CA). .. Semi-quantitative PCR was performed using a Peltier Thermal Cycler (PTC-200) using specific 3′and 5′ primers for GRP and final product was visualized on 1% agarose gel using a Gel Doc (Bio-Rad).

    Synthesized:

    Article Title: Overexpression of Cullin7 is associated with hepatocellular carcinoma progression and pathogenesis
    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted using TRIzol reagent (Invitrogen). .. As previously described [ ], First-strand cDNA was synthesized with a Super ScriptIII First-Strand Synthesis System (Invitrogen). qRT-PCR for cullin7 mRNA was performed on Applied Biosystems 7500 Fast Real-Time PCR system (Applied Biosystems) using One Step SYBR PrimeScript Plus RT-PCR kit (Takara Bio, Inc., Shiga, Japan) according to the manufacturer’s instruction.

    Isolation:

    Article Title: Antinociceptive Effect of Tetrandrine on LPS-Induced Hyperalgesia via the Inhibition of IKK? Phosphorylation and the COX-2/PGE2 Pathway in Mice
    Article Snippet: .. 2.9 Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was isolated from brain tissues and cultured cells using Trizol reagent, (Invitrogen, USA) according to the manufacturer's guidelines, followed by further purification using the RNeasy Mini Kits (Qiagen, USA). .. Purified RNA was reverse-transcribed into cDNA with random hexamers by the SuperScriptTM First-Strand Synthesis System kit (Qiagen, USA) and analyzed by real-time RT-PCR with the QuantiTect SYBR Green PCR Kits (Qiagen, USA), using a MJ DNA Engine Opticon2 qPCR System (MJ, USA).

    Article Title: Targeting Gastrin-Releasing Peptide Suppresses Neuroblastoma Progression via Upregulation of PTEN Signaling
    Article Snippet: .. Reverse transcription, semi-quantitative and quantitative real-time polymerase chain reaction (QRT-PCR) Total RNA was isolated using Trizol and reverse-transcribed to cDNA using High Capacity cDNA reverse transcription kit according to manufacturer's protocol (Applied Biosystems, Foster City, CA). .. Semi-quantitative PCR was performed using a Peltier Thermal Cycler (PTC-200) using specific 3′and 5′ primers for GRP and final product was visualized on 1% agarose gel using a Gel Doc (Bio-Rad).

    Article Title: HSP90 inhibition downregulates thymidylate synthase and sensitizes colorectal cancer cell lines to the effect of 5FU-based chemotherapy
    Article Snippet: .. Quantitative real time-polymerase chain reaction (qRT-PCR) Total RNA was isolated using TRIzol reagent (Invitrogen Corporation, Carlsbad, CA). .. The reverse transcription (RT) step was performed with MultiScribeTM reverse transcriptase (Applied Biosystems, USA).

    Article Title: Resveratrol Exerts Dosage-Dependent Effects on the Self-Renewal and Neural Differentiation of hUC-MSCs
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was isolated using Trizol reagent (Invitrogen) according to the instructions, and mRNA was reverse transcribed using PrimeScript™ RT-PCR Kit (TaKaRa, Japan). qRT-PCR were performed on an ABI 7500 real-time PCR system (Thermo Fisher Scientific, USA) using a SYBR Premix Ex Taq™ (Prefect Real Time, Takara, Japan). ..

    Article Title: Annexin A2 Coordinates STAT3 to Regulate the Invasion and Migration of Colorectal Cancer Cells In Vitro
    Article Snippet: .. RNA Extraction and Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA of tissue samples was extracted by using the RecoverAll™ Total Nucleic Acid Isolation Kit (Ambion, Austin, USA). .. RNeasy kits (Qiagen, Valencia, CA) were utilized to extract total mRNA from cells. qRT-PCR was performed on ABI 7500 with SYBR Premix Ex Taq™ Kit (Takara, Japan).

    Article Title: Doxycycline Attenuates Leptospira-Induced IL-1β by Suppressing NLRP3 Inflammasome Priming
    Article Snippet: .. Total RNA Isolation and Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA of cells and organs were extracted using TRIzol (Invitrogen, USA) following the manufacturer’s instructions. .. RNA was reverse transcribed into cDNA by using random primers from a TransScript One-Step gDNA Removal kit and cDNA Synthesis SuperMix (TransGen Biotech, China).

    Mouse Assay:

    Article Title: miR-22 inhibits mouse ovarian granulosa cell apoptosis by targeting SIRT1
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA were extracted from ovarian GCs of mice using a TRIZOL regent kit (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions. .. The purity of RNA was assessed by a NanoDrop 2000 UV-Vis spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA) at 260/280.

    Cell Culture:

    Article Title: Antinociceptive Effect of Tetrandrine on LPS-Induced Hyperalgesia via the Inhibition of IKK? Phosphorylation and the COX-2/PGE2 Pathway in Mice
    Article Snippet: .. 2.9 Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was isolated from brain tissues and cultured cells using Trizol reagent, (Invitrogen, USA) according to the manufacturer's guidelines, followed by further purification using the RNeasy Mini Kits (Qiagen, USA). .. Purified RNA was reverse-transcribed into cDNA with random hexamers by the SuperScriptTM First-Strand Synthesis System kit (Qiagen, USA) and analyzed by real-time RT-PCR with the QuantiTect SYBR Green PCR Kits (Qiagen, USA), using a MJ DNA Engine Opticon2 qPCR System (MJ, USA).

    Article Title: Exosomal miR-21-5p derived from gastric cancer promotes peritoneal metastasis via mesothelial-to-mesenchymal transition
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted from tissues, cultured cells and exosomes using the TRIzol reagent (Invitrogen) according to the manufacturer’s instructions. .. Then RNA was reverse transcribed into cDNA using PrimeScript RT Reagent (TaKaRa, Japan).

    Purification:

    Article Title: Antinociceptive Effect of Tetrandrine on LPS-Induced Hyperalgesia via the Inhibition of IKK? Phosphorylation and the COX-2/PGE2 Pathway in Mice
    Article Snippet: .. 2.9 Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was isolated from brain tissues and cultured cells using Trizol reagent, (Invitrogen, USA) according to the manufacturer's guidelines, followed by further purification using the RNeasy Mini Kits (Qiagen, USA). .. Purified RNA was reverse-transcribed into cDNA with random hexamers by the SuperScriptTM First-Strand Synthesis System kit (Qiagen, USA) and analyzed by real-time RT-PCR with the QuantiTect SYBR Green PCR Kits (Qiagen, USA), using a MJ DNA Engine Opticon2 qPCR System (MJ, USA).

    SYBR Green Assay:

    Article Title: Antinociceptive Effect of Tetrandrine on LPS-Induced Hyperalgesia via the Inhibition of IKK? Phosphorylation and the COX-2/PGE2 Pathway in Mice
    Article Snippet: 2.9 Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was isolated from brain tissues and cultured cells using Trizol reagent, (Invitrogen, USA) according to the manufacturer's guidelines, followed by further purification using the RNeasy Mini Kits (Qiagen, USA). .. Purified RNA was reverse-transcribed into cDNA with random hexamers by the SuperScriptTM First-Strand Synthesis System kit (Qiagen, USA) and analyzed by real-time RT-PCR with the QuantiTect SYBR Green PCR Kits (Qiagen, USA), using a MJ DNA Engine Opticon2 qPCR System (MJ, USA).

    Article Title: Strigolactone analogues induce apoptosis through activation of p38 and the stress response pathway in cancer cell lines and in conditionally reprogramed primary prostate cancer cells.
    Article Snippet: Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was extracted using Trizol (Invitrogen, Carlsbad, CA, USA) using the manufacturer’s protocol. .. PCR was performed in triplicate using an ABI-Prism 7900 instrument (Invitrogen, Foster City, CA) and SYBR Green (Invitrogen, Foster City, CA) according to the manufacturer’s protocol.

    Article Title: Doxycycline Attenuates Leptospira-Induced IL-1β by Suppressing NLRP3 Inflammasome Priming
    Article Snippet: Total RNA Isolation and Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA of cells and organs were extracted using TRIzol (Invitrogen, USA) following the manufacturer’s instructions. .. The qPCR reaction was performed using an Applied Bioscience 7500 thermocycler and FastStart Universal SYBR Green Master (Roche Applied Science, Germany).

    Article Title: Leucine-rich alpha-2-glycoprotein-1 is up-regulated in colorectal cancer and is a tumor promoter
    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted from tissues with TRIzol reagents (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s protocol. .. RT-PCR was performed with the SYBR green Premix Ex TaqII (Takara, Dalian, China) with Applied Bio-systems Step One Plus RT-PCR system (Applied Bio-systems, Carlsbad, CA, USA).

    CTG Assay:

    Article Title: Antinociceptive Effect of Tetrandrine on LPS-Induced Hyperalgesia via the Inhibition of IKK? Phosphorylation and the COX-2/PGE2 Pathway in Mice
    Article Snippet: 2.9 Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was isolated from brain tissues and cultured cells using Trizol reagent, (Invitrogen, USA) according to the manufacturer's guidelines, followed by further purification using the RNeasy Mini Kits (Qiagen, USA). .. The following oligonucleotide primers were used: COX-1 forward: 5′-TGC CCT CTG TAC CCA AAG AC-3′ , reverse: 5′-GGA CCC ATC TTT CCA GAG GT-3′ ; COX-2 forward: 5′-CGG AGA GAG TTC ATC CCT GA-3′ ; reverse: 5′-ATC CTT GAA AAG GCG CAG T-3′ ; GAPDH (control) forward: 5′-CTC ATG ACC ACA GTC CAT GC-3′ , reverse: 5′-CAC ATT GGG GGT AGG AAC AC-3′ .

    Incubation:

    Article Title: Targeting Gastrin-Releasing Peptide Suppresses Neuroblastoma Progression via Upregulation of PTEN Signaling
    Article Snippet: Reverse transcription, semi-quantitative and quantitative real-time polymerase chain reaction (QRT-PCR) Total RNA was isolated using Trizol and reverse-transcribed to cDNA using High Capacity cDNA reverse transcription kit according to manufacturer's protocol (Applied Biosystems, Foster City, CA). .. SsoFAST EvaGreen Supermix, cDNA and specific 3′ and 5′ primers were incubated together using the manufacturer's protocol (Bio-Rad).

    Polymerase Chain Reaction:

    Article Title: Antinociceptive Effect of Tetrandrine on LPS-Induced Hyperalgesia via the Inhibition of IKK? Phosphorylation and the COX-2/PGE2 Pathway in Mice
    Article Snippet: 2.9 Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was isolated from brain tissues and cultured cells using Trizol reagent, (Invitrogen, USA) according to the manufacturer's guidelines, followed by further purification using the RNeasy Mini Kits (Qiagen, USA). .. Purified RNA was reverse-transcribed into cDNA with random hexamers by the SuperScriptTM First-Strand Synthesis System kit (Qiagen, USA) and analyzed by real-time RT-PCR with the QuantiTect SYBR Green PCR Kits (Qiagen, USA), using a MJ DNA Engine Opticon2 qPCR System (MJ, USA).

    Article Title: Targeting Gastrin-Releasing Peptide Suppresses Neuroblastoma Progression via Upregulation of PTEN Signaling
    Article Snippet: Reverse transcription, semi-quantitative and quantitative real-time polymerase chain reaction (QRT-PCR) Total RNA was isolated using Trizol and reverse-transcribed to cDNA using High Capacity cDNA reverse transcription kit according to manufacturer's protocol (Applied Biosystems, Foster City, CA). .. Semi-quantitative PCR was performed using a Peltier Thermal Cycler (PTC-200) using specific 3′and 5′ primers for GRP and final product was visualized on 1% agarose gel using a Gel Doc (Bio-Rad).

    Article Title: Analysis of the miRNA Expression Profiles in the Zearalenone-Exposed TM3 Leydig Cell Line
    Article Snippet: Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was extracted using Trizol reagent (Invitrogen, Carlsbad, CA, USA) following the manufacturer’s procedure. .. Reverse transcription reactions were performed at the following parameters: 25 °C (mRNA) for 5 min, 50 °C for 15 min, and 85 °C for 5 min. PCR reactions were performed at the following parameters: 95 °C for 5 min followed by 40 cycles of 95 °C for 10 s and 60 °C for 30 s. A U6 small nuclear RNA was used as the endogenous control for the data normalization of miRNAs, and β-actin was then adopted as the internal control of mRNA expression.

    Article Title: Exosomal miR-21-5p derived from gastric cancer promotes peritoneal metastasis via mesothelial-to-mesenchymal transition
    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted from tissues, cultured cells and exosomes using the TRIzol reagent (Invitrogen) according to the manufacturer’s instructions. .. PCR reactions were performed using a 7500 Realtime PCR System (Applied Biosystems, Carlsbad, CA, USA).

    Article Title: Strigolactone analogues induce apoptosis through activation of p38 and the stress response pathway in cancer cell lines and in conditionally reprogramed primary prostate cancer cells.
    Article Snippet: Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was extracted using Trizol (Invitrogen, Carlsbad, CA, USA) using the manufacturer’s protocol. .. PCR was performed in triplicate using an ABI-Prism 7900 instrument (Invitrogen, Foster City, CA) and SYBR Green (Invitrogen, Foster City, CA) according to the manufacturer’s protocol.

    Article Title: miR-96 exerts carcinogenic effect by activating AKT/GSK-3β/β-catenin signaling pathway through targeting inhibition of FOXO1 in hepatocellular carcinoma
    Article Snippet: Quantitative real time polymerase chain reaction (qRT-PCR) Total RNA in tumor tissues and cells was obtained by Trizol kit (Thermo Fisher Scientific, Waltham, MA, USA) according to the the instruction manual. .. PCR reaction was performed according to the following reaction procedure: 95 °C for 20 s, 58 °C for 30 s and 74 °C for 30 s, with 40 cycles.

    Article Title: LINC00460 modulates KDM2A to promote cell proliferation and migration by targeting miR-342-3p in gastric cancer
    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted from tissues and cells using the Trizol reagent (Invitrogen) according to the manufacturer’s instructions. .. For microRNA analysis, qRT-PCR was performed using the TaqMan MicroRNA Reverse Transcription Kit, TaqMan Universal PCR Master Mix (Thermo Fisher Scientific), and the corresponding primers.

    Article Title: Leucine-rich alpha-2-glycoprotein-1 is up-regulated in colorectal cancer and is a tumor promoter
    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted from tissues with TRIzol reagents (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s protocol. .. Thereafter, 1 μg of RNA from each sample was reverse transcribed into complementary DNA (cDNA) using random primers with PCR machine (Thermo Fisher Scientific, Waltham, USA), and the cDNA was subjected to qRT-PCR for LRG1.

    Quantitative RT-PCR:

    Article Title: Antinociceptive Effect of Tetrandrine on LPS-Induced Hyperalgesia via the Inhibition of IKK? Phosphorylation and the COX-2/PGE2 Pathway in Mice
    Article Snippet: .. 2.9 Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was isolated from brain tissues and cultured cells using Trizol reagent, (Invitrogen, USA) according to the manufacturer's guidelines, followed by further purification using the RNeasy Mini Kits (Qiagen, USA). .. Purified RNA was reverse-transcribed into cDNA with random hexamers by the SuperScriptTM First-Strand Synthesis System kit (Qiagen, USA) and analyzed by real-time RT-PCR with the QuantiTect SYBR Green PCR Kits (Qiagen, USA), using a MJ DNA Engine Opticon2 qPCR System (MJ, USA).

    Article Title: Targeting Gastrin-Releasing Peptide Suppresses Neuroblastoma Progression via Upregulation of PTEN Signaling
    Article Snippet: .. Reverse transcription, semi-quantitative and quantitative real-time polymerase chain reaction (QRT-PCR) Total RNA was isolated using Trizol and reverse-transcribed to cDNA using High Capacity cDNA reverse transcription kit according to manufacturer's protocol (Applied Biosystems, Foster City, CA). .. Semi-quantitative PCR was performed using a Peltier Thermal Cycler (PTC-200) using specific 3′and 5′ primers for GRP and final product was visualized on 1% agarose gel using a Gel Doc (Bio-Rad).

    Article Title: Analysis of the miRNA Expression Profiles in the Zearalenone-Exposed TM3 Leydig Cell Line
    Article Snippet: .. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was extracted using Trizol reagent (Invitrogen, Carlsbad, CA, USA) following the manufacturer’s procedure. .. The total RNAs from each sample were reverse-transcribed to cDNA using the miRNA first-strand cDNA synthesis kit (by stem-loop) (Vazyme, Nanjing, China).

    Article Title: HSP90 inhibition downregulates thymidylate synthase and sensitizes colorectal cancer cell lines to the effect of 5FU-based chemotherapy
    Article Snippet: .. Quantitative real time-polymerase chain reaction (qRT-PCR) Total RNA was isolated using TRIzol reagent (Invitrogen Corporation, Carlsbad, CA). .. The reverse transcription (RT) step was performed with MultiScribeTM reverse transcriptase (Applied Biosystems, USA).

    Article Title: Resveratrol Exerts Dosage-Dependent Effects on the Self-Renewal and Neural Differentiation of hUC-MSCs
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was isolated using Trizol reagent (Invitrogen) according to the instructions, and mRNA was reverse transcribed using PrimeScript™ RT-PCR Kit (TaKaRa, Japan). qRT-PCR were performed on an ABI 7500 real-time PCR system (Thermo Fisher Scientific, USA) using a SYBR Premix Ex Taq™ (Prefect Real Time, Takara, Japan). ..

    Article Title: Annexin A2 Coordinates STAT3 to Regulate the Invasion and Migration of Colorectal Cancer Cells In Vitro
    Article Snippet: .. RNA Extraction and Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA of tissue samples was extracted by using the RecoverAll™ Total Nucleic Acid Isolation Kit (Ambion, Austin, USA). .. RNeasy kits (Qiagen, Valencia, CA) were utilized to extract total mRNA from cells. qRT-PCR was performed on ABI 7500 with SYBR Premix Ex Taq™ Kit (Takara, Japan).

    Article Title: Protein phosphatase 2A-B55δ enhances chemotherapy sensitivity of human hepatocellular carcinoma under the regulation of microRNA-133b
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted using TRIzol® Reagent (Ambion, TX, USA) and reverse-transcribed into cDNA using PrimeScript™ RT reagent kit (TaKaRa, Otsu, Japan). qRT-PCR was performed with SYBR® Premix Ex Taq ™ II kit (TaKaRa) in a CFX96 Touch™ Real-Time PCR Detection System (Bio-Rad, CA, USA) using the primers (Sangon Biotech, Shanghai, China) shown in Additional file : Table S1. ..

    Article Title: Exosomal miR-21-5p derived from gastric cancer promotes peritoneal metastasis via mesothelial-to-mesenchymal transition
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted from tissues, cultured cells and exosomes using the TRIzol reagent (Invitrogen) according to the manufacturer’s instructions. .. Then RNA was reverse transcribed into cDNA using PrimeScript RT Reagent (TaKaRa, Japan).

    Article Title: Strigolactone analogues induce apoptosis through activation of p38 and the stress response pathway in cancer cell lines and in conditionally reprogramed primary prostate cancer cells.
    Article Snippet: .. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was extracted using Trizol (Invitrogen, Carlsbad, CA, USA) using the manufacturer’s protocol. .. 1μg RNA was reverse-transcribed in a total volume of 20 μl using the High Capacity cDNA kit (Invitrogen).

    Article Title: miR-96 exerts carcinogenic effect by activating AKT/GSK-3β/β-catenin signaling pathway through targeting inhibition of FOXO1 in hepatocellular carcinoma
    Article Snippet: .. Quantitative real time polymerase chain reaction (qRT-PCR) Total RNA in tumor tissues and cells was obtained by Trizol kit (Thermo Fisher Scientific, Waltham, MA, USA) according to the the instruction manual. ..

    Article Title: Overexpression of Cullin7 is associated with hepatocellular carcinoma progression and pathogenesis
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted using TRIzol reagent (Invitrogen). .. As previously described [ ], First-strand cDNA was synthesized with a Super ScriptIII First-Strand Synthesis System (Invitrogen). qRT-PCR for cullin7 mRNA was performed on Applied Biosystems 7500 Fast Real-Time PCR system (Applied Biosystems) using One Step SYBR PrimeScript Plus RT-PCR kit (Takara Bio, Inc., Shiga, Japan) according to the manufacturer’s instruction.

    Article Title: LINC00460 modulates KDM2A to promote cell proliferation and migration by targeting miR-342-3p in gastric cancer
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted from tissues and cells using the Trizol reagent (Invitrogen) according to the manufacturer’s instructions. .. For microRNA analysis, qRT-PCR was performed using the TaqMan MicroRNA Reverse Transcription Kit, TaqMan Universal PCR Master Mix (Thermo Fisher Scientific), and the corresponding primers.

    Article Title: Doxycycline Attenuates Leptospira-Induced IL-1β by Suppressing NLRP3 Inflammasome Priming
    Article Snippet: .. Total RNA Isolation and Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA of cells and organs were extracted using TRIzol (Invitrogen, USA) following the manufacturer’s instructions. .. RNA was reverse transcribed into cDNA by using random primers from a TransScript One-Step gDNA Removal kit and cDNA Synthesis SuperMix (TransGen Biotech, China).

    Article Title: miR-22 inhibits mouse ovarian granulosa cell apoptosis by targeting SIRT1
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA were extracted from ovarian GCs of mice using a TRIZOL regent kit (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions. .. The purity of RNA was assessed by a NanoDrop 2000 UV-Vis spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA) at 260/280.

    Expressing:

    Article Title: Analysis of the miRNA Expression Profiles in the Zearalenone-Exposed TM3 Leydig Cell Line
    Article Snippet: Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was extracted using Trizol reagent (Invitrogen, Carlsbad, CA, USA) following the manufacturer’s procedure. .. Reverse transcription reactions were performed at the following parameters: 25 °C (mRNA) for 5 min, 50 °C for 15 min, and 85 °C for 5 min. PCR reactions were performed at the following parameters: 95 °C for 5 min followed by 40 cycles of 95 °C for 10 s and 60 °C for 30 s. A U6 small nuclear RNA was used as the endogenous control for the data normalization of miRNAs, and β-actin was then adopted as the internal control of mRNA expression.

    Article Title: Resveratrol Exerts Dosage-Dependent Effects on the Self-Renewal and Neural Differentiation of hUC-MSCs
    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was isolated using Trizol reagent (Invitrogen) according to the instructions, and mRNA was reverse transcribed using PrimeScript™ RT-PCR Kit (TaKaRa, Japan). qRT-PCR were performed on an ABI 7500 real-time PCR system (Thermo Fisher Scientific, USA) using a SYBR Premix Ex Taq™ (Prefect Real Time, Takara, Japan). .. Relative expression levels of different genes were calculated using the 2−ΔΔCt .

    Article Title: Protein phosphatase 2A-B55δ enhances chemotherapy sensitivity of human hepatocellular carcinoma under the regulation of microRNA-133b
    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted using TRIzol® Reagent (Ambion, TX, USA) and reverse-transcribed into cDNA using PrimeScript™ RT reagent kit (TaKaRa, Otsu, Japan). qRT-PCR was performed with SYBR® Premix Ex Taq ™ II kit (TaKaRa) in a CFX96 Touch™ Real-Time PCR Detection System (Bio-Rad, CA, USA) using the primers (Sangon Biotech, Shanghai, China) shown in Additional file : Table S1. .. Quantitative analysis of miRNA expression was performed with the Bulge-Loop™ hsa-miR-133b qRT-PCR primer set (Ribobio, Guangzhou, China).

    Article Title: LINC00460 modulates KDM2A to promote cell proliferation and migration by targeting miR-342-3p in gastric cancer
    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted from tissues and cells using the Trizol reagent (Invitrogen) according to the manufacturer’s instructions. .. For mRNA analysis, qRT-PCR was performed using the TaqMan High-Capacity cDNA Reverse Transcription Kit, TaqMan Fast PCR Master Mix (Thermo Fisher Scientific) and the corresponding primers. β-actin was used as an internal control to normalize KDM2A expression. qRT-PCR was performed in triplicate on a RealPlex4 real-time PCR detection system from Eppendorf Co. Ltd (Hamburg, Germany).

    Article Title: miR-22 inhibits mouse ovarian granulosa cell apoptosis by targeting SIRT1
    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA were extracted from ovarian GCs of mice using a TRIZOL regent kit (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions. .. Total RNA was reversely transcribed using TaqMan miRNA reverse transcription kit (Applied Biosystems Inc., Foster City, CA, USA) to detect the expression level of miR-22.

    Article Title: Leucine-rich alpha-2-glycoprotein-1 is up-regulated in colorectal cancer and is a tumor promoter
    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted from tissues with TRIzol reagents (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s protocol. .. Actin was used as the endogenous control for detection of LRG1 mRNA expression.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Resveratrol Exerts Dosage-Dependent Effects on the Self-Renewal and Neural Differentiation of hUC-MSCs
    Article Snippet: .. Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was isolated using Trizol reagent (Invitrogen) according to the instructions, and mRNA was reverse transcribed using PrimeScript™ RT-PCR Kit (TaKaRa, Japan). qRT-PCR were performed on an ABI 7500 real-time PCR system (Thermo Fisher Scientific, USA) using a SYBR Premix Ex Taq™ (Prefect Real Time, Takara, Japan). ..

    Article Title: Overexpression of Cullin7 is associated with hepatocellular carcinoma progression and pathogenesis
    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted using TRIzol reagent (Invitrogen). .. As previously described [ ], First-strand cDNA was synthesized with a Super ScriptIII First-Strand Synthesis System (Invitrogen). qRT-PCR for cullin7 mRNA was performed on Applied Biosystems 7500 Fast Real-Time PCR system (Applied Biosystems) using One Step SYBR PrimeScript Plus RT-PCR kit (Takara Bio, Inc., Shiga, Japan) according to the manufacturer’s instruction.

    Article Title: Leucine-rich alpha-2-glycoprotein-1 is up-regulated in colorectal cancer and is a tumor promoter
    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted from tissues with TRIzol reagents (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s protocol. .. RT-PCR was performed with the SYBR green Premix Ex TaqII (Takara, Dalian, China) with Applied Bio-systems Step One Plus RT-PCR system (Applied Bio-systems, Carlsbad, CA, USA).

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Antinociceptive Effect of Tetrandrine on LPS-Induced Hyperalgesia via the Inhibition of IKK? Phosphorylation and the COX-2/PGE2 Pathway in Mice
    Article Snippet: 2.9 Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was isolated from brain tissues and cultured cells using Trizol reagent, (Invitrogen, USA) according to the manufacturer's guidelines, followed by further purification using the RNeasy Mini Kits (Qiagen, USA). .. The following oligonucleotide primers were used: COX-1 forward: 5′-TGC CCT CTG TAC CCA AAG AC-3′ , reverse: 5′-GGA CCC ATC TTT CCA GAG GT-3′ ; COX-2 forward: 5′-CGG AGA GAG TTC ATC CCT GA-3′ ; reverse: 5′-ATC CTT GAA AAG GCG CAG T-3′ ; GAPDH (control) forward: 5′-CTC ATG ACC ACA GTC CAT GC-3′ , reverse: 5′-CAC ATT GGG GGT AGG AAC AC-3′ .

    Spectrophotometry:

    Article Title: miR-22 inhibits mouse ovarian granulosa cell apoptosis by targeting SIRT1
    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA were extracted from ovarian GCs of mice using a TRIZOL regent kit (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions. .. The purity of RNA was assessed by a NanoDrop 2000 UV-Vis spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA) at 260/280.

    Software:

    Article Title: Strigolactone analogues induce apoptosis through activation of p38 and the stress response pathway in cancer cell lines and in conditionally reprogramed primary prostate cancer cells.
    Article Snippet: Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Total RNA was extracted using Trizol (Invitrogen, Carlsbad, CA, USA) using the manufacturer’s protocol. .. Primers were designed using PrimerQuest software (Integrated DNA Technologies).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher quantitative real time polymerase chain reaction qrt pcr total rna
    Up-regulation of miR-96 and down-regulation of FOXO1 in tumor tissues and HepG2 cells. a miR-96 relative expression in tumor tissues and non-tumor tissues by <t>qRT-PCR;</t> b FOXO1 mRNA relative expression in tumor tissues and non-tumor tissues by qRT-PCR; c FOXO1 protein relative expression in tumor tissues and non-tumor tissues by Western blot; d miR-96 relative expression in L02 and HepG2 cells by qRT-PCR; e FOXO1 mRNA relative expression in L02 and HepG2 cells by qRT-PCR; f FOXO1 protein relative expression in L02 and HepG2 cells by Western blot. ** P
    Quantitative Real Time Polymerase Chain Reaction Qrt Pcr Total Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 364 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time polymerase chain reaction qrt pcr total rna/product/Thermo Fisher
    Average 99 stars, based on 364 article reviews
    Price from $9.99 to $1999.99
    quantitative real time polymerase chain reaction qrt pcr total rna - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    Image Search Results


    Up-regulation of miR-96 and down-regulation of FOXO1 in tumor tissues and HepG2 cells. a miR-96 relative expression in tumor tissues and non-tumor tissues by qRT-PCR; b FOXO1 mRNA relative expression in tumor tissues and non-tumor tissues by qRT-PCR; c FOXO1 protein relative expression in tumor tissues and non-tumor tissues by Western blot; d miR-96 relative expression in L02 and HepG2 cells by qRT-PCR; e FOXO1 mRNA relative expression in L02 and HepG2 cells by qRT-PCR; f FOXO1 protein relative expression in L02 and HepG2 cells by Western blot. ** P

    Journal: Cancer Cell International

    Article Title: miR-96 exerts carcinogenic effect by activating AKT/GSK-3β/β-catenin signaling pathway through targeting inhibition of FOXO1 in hepatocellular carcinoma

    doi: 10.1186/s12935-019-0756-7

    Figure Lengend Snippet: Up-regulation of miR-96 and down-regulation of FOXO1 in tumor tissues and HepG2 cells. a miR-96 relative expression in tumor tissues and non-tumor tissues by qRT-PCR; b FOXO1 mRNA relative expression in tumor tissues and non-tumor tissues by qRT-PCR; c FOXO1 protein relative expression in tumor tissues and non-tumor tissues by Western blot; d miR-96 relative expression in L02 and HepG2 cells by qRT-PCR; e FOXO1 mRNA relative expression in L02 and HepG2 cells by qRT-PCR; f FOXO1 protein relative expression in L02 and HepG2 cells by Western blot. ** P

    Article Snippet: Quantitative real time polymerase chain reaction (qRT-PCR) Total RNA in tumor tissues and cells was obtained by Trizol kit (Thermo Fisher Scientific, Waltham, MA, USA) according to the the instruction manual.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot

    FOXO1 was directly suppressed by miR-96. a Prediction of binding sites for FOXO1 and miR-96 by Target Scan; b Luciferase reporter assay; c FOXO1 mRNA expression by qRT-PCR; d FOXO1 protein expression by Western blot. ## P

    Journal: Cancer Cell International

    Article Title: miR-96 exerts carcinogenic effect by activating AKT/GSK-3β/β-catenin signaling pathway through targeting inhibition of FOXO1 in hepatocellular carcinoma

    doi: 10.1186/s12935-019-0756-7

    Figure Lengend Snippet: FOXO1 was directly suppressed by miR-96. a Prediction of binding sites for FOXO1 and miR-96 by Target Scan; b Luciferase reporter assay; c FOXO1 mRNA expression by qRT-PCR; d FOXO1 protein expression by Western blot. ## P

    Article Snippet: Quantitative real time polymerase chain reaction (qRT-PCR) Total RNA in tumor tissues and cells was obtained by Trizol kit (Thermo Fisher Scientific, Waltham, MA, USA) according to the the instruction manual.

    Techniques: Binding Assay, Luciferase, Reporter Assay, Expressing, Quantitative RT-PCR, Western Blot

    The mRNA expression of HDAC9 measured by qRT-PCR. Notes: ( A ) HDAC9 expression in breast cancer tissues and paired normal tissues. The expression of HDAC9 was higher in breast cancer tissues than that in the matched normal tissues (*** P

    Journal: OncoTargets and therapy

    Article Title: Overexpression of HDAC9 is associated with poor prognosis and tumor progression of breast cancer in Chinese females

    doi: 10.2147/OTT.S164583

    Figure Lengend Snippet: The mRNA expression of HDAC9 measured by qRT-PCR. Notes: ( A ) HDAC9 expression in breast cancer tissues and paired normal tissues. The expression of HDAC9 was higher in breast cancer tissues than that in the matched normal tissues (*** P

    Article Snippet: RNA extraction and quantitative real-time polymerase chain reaction (qRT-PCR) Total RNA was extracted from the tissues and cells using TRIzol reagent (Thermo Fisher Scientific) as per the manufacturer’s instructions.

    Techniques: Expressing, Quantitative RT-PCR