Fig. 6: IAA94 prevents neuronal death induced by oxidative stress.

a–c Cell viability of primary cortical neurons was measured by Trypan blue staining and CCK-8. At DIV8, neurons were treated with or without 300 μM H₂O₂ for 6 h in the presence or absence of 50 μM IAA94. Then cell viability was determined via Trypan blue staining (by the ratio of the unstained cell number to the total cell number; about 100 cells per coverslip were counted and at least 3 coverslips were used) or CCK-8 assay (n = 3 experiments). Scale bar = 100 μm. d, e Cleaved caspase3 levels in neurons treated with or without 300 μM H₂O₂ for 6 h in the presence or absence of 50 μM IAA94. Cell lysates were subjected to immunostaining analysis. Relative cleaved caspase3 levels were quantified in E (n = 3 experiments). f, h Protein levels of CLIC4 and cleaved caspase3 in hippocampi of mice injected with indicated drugs. Adult mice were injected with 10 μl 50 mM IAA94 into lateral ventricle, then 1 h later, 1 μl KA (1 μg/μl) or 1 μl saline was injected into the same location. 24 h after injection, mouse hippocampi were collected and lysed for immunoblotting. Five mice were use in each group. Relative CLIC4 and cleaved caspase3 levels were quantified in g and h (n = 3 experiments). Data are presented as the mean and SEM, and were analyzed by one-way ANOVA test followed by Tukey test. *P < 0.05; **P < 0.01; n.s. not significant