Fig. 3.

Alterations in the genetic pathways controlling alcohol metabolism, oxidative stress response and the control of DNA methylation. Murine embryonic stem cells were cultured in varying concentrations of ethanol (80 mg/dL, 160 mg/dL, or 240 mg/dL) for four days, followed by a no-ethanol recovery period for ten days. Samples were taken at Day-4, Day-8, and Day-14, and transcripts encoding genes involved in A) alcohol metabolism and the oxidative stress response, B) H3K27, H3K9 and DNA methyltransferase enzymes, C) TET family of Fe(II) and a-KG- dependent dioxygenases, and D) histone demethylases examined using RT-qPCR. Ct values were normalized to the geometric mean of Gapdh, Hprt, and Ppia and graphed relative to the control treatment. Graphs represent three independent replicates (N = 3), with two independent RT reactions and three qPCR measurements for each RT. Differences were measured using a one-way ANOVA, error bars represent SEM. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.