Figure 8.

FoxO3a silencing reduces hypoxic Bnip3 expression and mitochondrial damage. Cortical neuron cultures were treated with lentiviral particles expressing shRNA to reduce the transcription of FoxO3a. A, B, Real-time PCR indicated a dramatic reduction of FoxO3a mRNA levels (A) and immunoreactivity (B). C, Hypoxia (24 h) enhanced Bnip3 mRNA levels in a manner significantly attenuated by shRNA for FoxO3a. D, E, FoxO3a shRNA significantly restored the ratio of JC-1 aggregate (red) to monomer (green) in hypoxia, indicating that FoxO3a contributed to hypoxic mitochondrial depolarization. F, G, FoxO3a shRNA significantly reduced hypoxic neuron death. A and B: *p < 0.05, compared with control shRNA group using t test. C, E, and G: **p < 0.01, compared with normoxic control; and ††p < 0.01, compared with the hypoxic group using ANOVA with Student–Newman–Keuls test. All data are reported as the mean ± SEM (n = 5–7). Scale bars: D, 25 μm; F, 40 μm.