Acetylation enhances TET2 function in protecting against abnormal DNA methylation during oxidative stress [shTET2 - H2O2 - methylation]

Genome wide DNA methylation profilings (Bisulfite (BS), or oxidative bisulfite (oxBS)) were done for A2780 cells 1) infected with control virus, no H2O2 (Scr_mock) treatment, 2) infected with control virus with H2O2 treatment (30 min plus 2.5 h resting) (Scr_H2O2), and 3) infected with shTET2 virus, with H2O2 treatment (30 min plus 2.5 h resting) (shTET2_H2O2). Each genomic DNA was splitted equally to two aliquots. One aliquote was subjected to oxidation (oxBS) and one to mock oxidation (BS) prior to bisulfite treatment (CEGX protocol). The Illumina’s Infinium Human Methylation450 Beadchip Kit (WG-314-1001) was used to obtain DNA methylation profiles across approximately 450,000 CpGs. 5hmC levels were calculated by subtracting oxBS values from BS values. Genome wide DNA methylation profilings (Bisulfite (BS), or oxidative bisulfite (oxBS)) were done for A2780 cells 1) infected with control virus, no H2O2 (Scr_mock) treatment, 2) infected with control virus with H2O2 treatment (30 min plus 2.5 h resting) (Scr_H2O2), and 3) infected with shTET2 virus, with H2O2 treatment (30 min plus 2.5 h resting) (shTET2_H2O2).