Figure 9.

Inhibition of autophagy by 3-methyladenine (3-MA) exacerbates the injury of primary SMNs exposed to OGD. (A) Primary SMNs in normal conditions were treated with different dosages of 3-MA to select its safe dosage. (B) The autophagy inhibition effect of 5 mM 3-MA was confirmed by Western blot. To test it simple and clear, SMNs were subjected to OGD for 5 h in the presence or absence of 3-MA, and their expressions of LC3 I and LC3 II were detected. Results indicated that this dosage can significantly block autophagy in SMNs under OGD conditions. (C) The dosage 5 mM was applied in all the time points, and the cell viability of primary SMNs with or without 3-MA was assessed at different time points using the CCK8 kit. Results were expressed as the percentage of CCK8 reduction, and the absorbance of the control cells was set at 100%. Data were presented as mean ± SD (n = 5 independent experiments). Statistical comparisons were carried out with one-way ANOVA (A) or two-way ANOVA (C). ^#p < 0.05 compared with the control group. *p < 0.05 compared with cell viability in the same time point under OGD conditions.