Fig. 6

Hydrogen peroxide significantly alters ARNT2 protein expression. Four doses were applied: a–d 25, 50, 100, and 300 μM H₂O₂. Graphs represent the ARNT2/β-actin ratios of protein expression from western blotting. Points represent six biological replicates, with two technical replicates each, where lines and whiskers indicate mean and standard deviation, with representative blots. Cytotoxicity measured by lactic dehydrogenase (LDH) release is reported as the % cytotoxicity or % of total death relative to wells lysed with lysis buffer. Supernatants were collected at intervals following H₂O₂ treatment and compared to spontaneous LDH release in untreated controls. p ≤ 0.05*, p ≤ 0.01**, p ≤ 0.001***, repeated measures ANOVA on ranks with Tukey’s multiple comparison test to unstimulated. e Representative phase contrast photomicrographs show neuronal bodies with numerous complex dendrites and processes throughout the culture in untreated cultures. Addition of increasing doses of H₂O₂ over time leads to a loss in cell viability evidenced by decreasing processes and complexity of neurons with increasing amounts of cellular debris and condensed nuclei (asterisk)