Fig. 2.

Hypoxia-induced EZH2 regulated TGFBR2 promoter hypermethylation and contributed to its epigenetic silencing in PCa. a. TGFBR2 expression in PCa cell lines elevated significantly after the 5-Aza-DC treatment via RT-qPCR. b. Graphic presentation of putative CpG island in promoter of TGFBR2 and primer for methylation specific PCR, as designed by MethPrimer website. c. Methylation specific PCR detected the methylation status of TGFBR2 promoter in six pairs of prostate cancer samples (T1-T6) and corresponding noncancerous tissues (N1-N6). M, methylation primer; U, unmethylation primer. d. Significant negative correlation between the expression of EZH2 and TGFBR2 via data from TCGA. e-f. The expression levels of TGFBR2 elevated significantly after treatment with siRNAs targeting EZH2 or EZH2 inhibitor DZNep, as detected by RT-qPCR (E) and western blot (F). G. Methylation specific PCR results showed DZNep and BIX-01294 partially decreased the methylation levels of PCa cell lines. h-i. Hypoxia induced both the mRNA (H) and protein expression (I) of EZH2.**, P < 0.01