Fig 1: The regulatory effect of REV1 on SERTAD2 is partially dependent on Rad18.A Proteins from REV1-silenced A549 and H1299 cells were harvested and examined by Western blot (n = 3). B Scatter plots of SERTAD2 expression vs. Rad18, PCNA, and RPA32 expression in lung adenocarcinoma samples acquired from the GEPIA database. C The protein level of SERTAD2 was decreased in Rad18-depleted cells (n = 3). D A549 and H1299 cells transfected with SiRad18 and Flag-REV1 were harvested and examined by Western blot (n = 3). E Cells transfected with SiRad18 and Flag-REV1 were harvested for colony formation assays. Representative pictures of the indicated groups and quantitative analysis of the colony counts are shown. ***P < 0.001 (n = 3). F EdU incorporation assays were used to test the proliferation of cells transfected with SiRad18 and Flag-REV1. Representative pictures of the indicated groups and statistical analysis results are presented. *P < 0.05, **P < 0.01, ***P < 0.001 (n = 3). Scale bar: 50 µm. G–I A549 and H1299 cells transfected with SiRad18 and SFB-SERTAD2 were harvested for cell proliferation assays. Representative pictures of the indicated groups and the statistical analysis results are shown. **P < 0.01, ***P < 0.001 (n = 3). Scale bar: 50 µm.
Fig 2: REV1 regulates the expression of SERTAD2 in lung cancer cells.A Volcano plots showing differentially expressed genes in REV1-depleted vs. scramble siRNA-transfected A549 cells. B Venn diagram showing the number of genes in each indicated set. C Clustering heatmap of the ten most deregulated genes affected by REV1 silencing. D The mRNA levels of the indicated molecules in scramble siRNA-transfected and REV1-silenced cells were measured by PCR. **P < 0.01, ***P < 0.001, n.s. P > 0.05 (n = 4). E The mRNA level of the indicated molecules in DMSO-treated and JH-RE-06-treated cells was examined by PCR. *P < 0.05, **P < 0.01, ***P < 0.001, n.s. P > 0.05 (n = 4). F The protein level of the indicated molecules in scramble siRNA-transfected and REV1-silenced cells was measured by Western Blot (n = 3). G The protein level of the indicated molecules in DMSO-treated and JH-RE-06-treated cells was examined by Western Blot (n = 3). H Scatter plot showing the correlation between REV1 and SERTAD2 expression in lung adenocarcinoma samples available in the GEPIA database. I Representative images of IHC staining for SERTAD2 in DMSO-treated and JH-RE-06-treated xenograft tumors and statistical histograms of the IHC score. **P < 0.01, n.s. P > 0.05 (n = 3). Scale bar: 50 µm.
Fig 3: The inhibitory effect of REV1 silencing on the proliferation of lung cancer cells is partially dependent on SERTAD2.A A549 and H1299 cells transfected with the indicated siRNAs and SFB-SERTAD2 were harvested and analyzed by Western blot (n = 3). B Cells transfected with the indicated siRNAs and SFB-SERTAD2 were seeded in a six-well plate in triplicate and grown for two weeks. Representative pictures of colony formation in each group and quantitative analysis of the colony counts are shown. **P < 0.01, ***P < 0.001 (n = 3). C EdU incorporation assays were used to test the proliferation of cells transfected with the indicated siRNAs and SFB-SERTAD2. **P < 0.01, ***P < 0.001 (n = 3). Scale bar: 50 µm. D A549 and H1299 cells transfected with the indicated siRNAs were harvested and analyzed by Western blot (n = 3). E Colony formation assays were conducted to examine the proliferation ability of cells transfected with the indicated siRNAs. ***P < 0.001, n.s. P > 0.05 (n = 3). F Representative fluorescence images of each group transfected with the indicated siRNAs from EdU incorporation assays and statistical analysis results are presented. ***P < 0.001, n.s. P > 0.05 (n = 3). Scale bar: 50 µm.
Supplier Page from Abcam for Anti-TRIP-Br2 antibody