Figure 5. Validation of methylation changes by deep bisulfite sequencing.

Schematic outline of the promoter regions of (A) KRT75 (B) SEC31L2, (C) DDAH2, and (D) TET2. Vertical lines represent individual CpG dinucleotides, blue arrowheads indicate CpG markers represented on the array. PCR amplification was performed on equimolar sample pools. PCR amplicons for sequencing are shown as grey horizontal bars, sequencing results are shown as heatmaps. Each row represents one sequence read, individual red boxes represent methylated CpG dinucleotides, green boxes represent unmethylated CpG dinucleotides, sequencing gaps are shown in white. Sequencing coverage ranged from 31x to 172x, as indicated. The deamination efficiency was >99% for all samples analyzed.