Fig. 9.

Inhibition of oxidative stress and cell cycle events alleviates shMettl3-induced cell cycle abnormalities and neurodegenerative changes in primary neurons. Primary cortical neurons were infected by AAV-GFP-shRNA at around DIV7. About 8–9 days after infection, primary neurons were used for analysis. Representative immunoblot (A) and quantitative analysis of CCND1 (B), CCND2 (C), CCNB1 (D) and cleaved caspase 3 (E) in primary neurons after AAV-shRNAs infection with/without co-treatment of 100 μM NAC or 40 nM flavopiridol. Representative immunofluorescence image (F) and quantitative analysis of PSD95 (G) and MAP 2 (H) in the neurites of primary cortical neurons after AAV-shRNAs infection with/without co-treatment of 100 μM NAC or 40 nM flavopiridol. (I) Cell death was measured by propidium iodide (PI) uptake in primary cortical neurons after AAV-shRNA infection with/without co-treatment of 100 μM NAC or 40 nM flavopiridol (Data are means±SEM from at least 3 independent experiments, *p < 0.05, **p < 0.01, ***p < 0.001; B-E, G-I one-way ANOVA with bonferroni’s correction)