Figure 3.

Application of four-plex MS-NaME (ATM, RARb2, MGMT and GSTP1 gene promoters) on bisulfite-treated DNA to enrich serially diluted levels of methylated or unmethylated DNA. 0.1–10% methylated DNA or 0.1–10% unmethylated DNA were treated with U probes and M probes, respectively. (A and B) ATM and RARb2 were assessed via digital Methylight to determine changes in methylation and unmethylation levels as compared to no treatment. Error bars represent the standard error of the mean from three independent experiments. (C and D): Bisulfite Sanger sequencing for the four genes was conducted with 0.1% methylated DNA or 0.1% unmethylated DNA following MS-NaME treatment with U probes and M probes, respectively.