To determine if EtOH-induced hypermethylation could account for decreases in gene expression we examined the effect of 5-azacytidine (1 μM) on the expression level of genes that were downregulated upon EtOH treatment (Fig. 4H). We chose four genes (ACADY4, FGF17, HOXA1 and PHOSPHO1) out of top ten genes from combined analysis that were identified as most significantly altered. Co-administration of EtOH with this demethylating agent overcame the gene silencing effect of EtOH (Fig. 4H), suggesting that the changes in level of these genes are mostly due to DNA methylation events.
