Fig. 4.

The effect of Mn on autophagy. Short- (4, 8, and 12 h) and long-term (1, 7, 14, and 28 days) after Mn injection, TEM and Western blotting were conducted to evaluate autophagy. a TEM observation on increased autophagy (a control; b, c, d 4, 8, and 12 h post Mn injection) and dysfunctional lysosomes (e–h 1, 7, 14, and 28 days post Mn injection). Figure 4 A, b–d shows that the autophagosomes-containing double membrane increased compared with that of control, Fig. 4 A, e–h shows that increased dysfunctional lysosomes-containing dense granules compared with the control. b, e Effect of Mn on protein expression of Beclin 1, LC3 II, and LC3 I. c, d, f, g Densitometry analysis of Beclin 1 protein levels and LC3 II levels relative to LC3 I was performed using three independent experiments. β-Actin was used as control for protein loading. Above results were obtained after three independent experiments (mean ± SD; one-way ANOVA with Newman–Keuls post hoc analysis, *p < 0.05 and **p < 0.01)