Global DNA methylation in brain regions of adult animals. Global methylation was analyzed using Methylamp kit (Epigentek) in animals from control (gray bars) or HF-fed (black bars) dams. Values are mean ± sem. Two-way ANOVA revealed a main effect for group (*, P < 0.0001), with offspring from HF-fed dams demonstrating a significant reduction in global DNA methylation. Furthermore, global methylation levels in the VTA were significantly lower than in all other brain regions (a, P < 0.008), and global methylation in PFC was significantly lower than in the hypothalamus (b, P < 0.008) (n = 5 per group).