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. Author manuscript; available in PMC: 2018 Dec 16.
Published in final edited form as: Chem Res Toxicol. 2018 Jan 8;31(2):88–96. doi: 10.1021/acs.chemrestox.7b00248

Figure 5.

Figure 5

Methylation ratio determined by BGS and MeDIP of the first 5 CpGs from −1226 to −1086 upstream the TSS of the Nrf2 promoter in TRAMP C1 cells after 3 days of treatment with 0.1% DMSO (control), mixture of 5-aza (500 nM) and TSA (100 nM), E10 (100 nM) or F10 (100 nM). The data are indicated as the means ± SD from 3 independent tests.*. p < 0.05 and **, p < 0.01 compared with the control group. BGS assay in evaluating the influence of E10 (100 nM) and F10 (100 nM) compared with the control (0.1% DMSO), 5-aza (500 nM) and TSA (100 nM) on methylation of the Nrf2 promoter. Solid circles denote methylated CpGs, while hollow circles unmethylated ones (Figure 5A). MeDIP assay in examining the effects of E10 (100 nM) and F10 (100 nM) compared with the control (0.1% DMSO), 5- aza (500 nM) and TSA (100 nM) on Nrf2 promoter methylation status. qPCR was utilized to quantify the amount of MeDIP DNAs relative to their inputs by calculating the standard curve from a serial dilution of their input DNAs. The relative methylated DNA ratio was compared with the control (Figure 5B).