Fig. 8
Gene regulation by DNMT3a and TET2 occurs in promoter DNA methylation -dependent and independent manners in sorafenib cells. (a), qPCR measuring the indicated TSG expression in SorafenibR Hep3b and Huh7 cells. (b), qPCR measuring the expression of p15 and SOCS2 in SorafenibR Hep3b cells either infected with DNMT3a or/and TET2 virus vectors or treated with bobcat339 for 48 h. (c), DNA (1 µg) from parental or SorafenibR Hep3b cells treated or untreated with 2 μm decitabine for 48 h were digested by HpaII or BstuI, and PCR was performed using primers specific for p15 or SOCS2 gene promoter. HpaII indicates no digestion, then hypermethylated; BstUI, no digestion, then hypomethylated. (d), MeDIP assays were performed in SorafenibR and parental Hep3b cells, and the 5mC-enriched DNA was subjected to qPCR using primers of p15 or SOCS2 gene promoter. (e), ChIP assays using antibodies for HDAC2, DNMT3a and TET2 were performed in SorafenibR Hep3b cells, and the ChIP-enriched DNA was subjected to qPCR using primers specific for p15 or SOCS2 promoter. (f), qPCR to measure the expression of CDK1, CCNA2 and RASEF in SorafenibR Hep3b and Huh7 cells. (g), qPCR to measure the expression of CDK1, CCNA2 and RASEF in SorafenibR Hep3b cells either infected with DNMT3a or/and TET2 virus vector or treated with bobcat339 for 48 h. Data represents three independent experiments. *P < 0.05; ns, not significant. SR, SorafenibR; Scr, scramble; Par, parental; Deci, decitabine