Maternal diabetes and high glucose in vitro induce H2A.X phosphorylation. A. Phosphorylation of H2A.X in E8.75 embryos from nondiabetic or diabetic dams, in the presence or absence of SOD1 overexpression. B. Phosphorylation of H2A.X in neural stem cells treated with glucose (5, 16.7, 25, 33.3 mM) for 48 hours. C. Phosphorylation of H2A.X in neural stem cells treated with normal glucose (5 mM) plus mannitol (0, 11.7, 20, 18.3 mM) for 48 hours. D. Phosphorylation of H2A.X in neural stem cells treated with normal (5 mM) or high glucose (25 mM) for 48 hours, in the presence or absence of Tempol (100 μM). Within each panel, the membrane was probed first for the phosphorylated protein and sequentially stripped for probing of the total protein content and β-actin. The quantification of the blot is shown in the bar graph. Experiments were repeated three times (n = 3). Values are the means ± SEM from three separate experiments. * indicates significant differences (P < 0.05) compared to the other three groups (A, D) or the normal glucose (5 mM) groups (B).