Fig. 8

ZnO NPs regulates autophagic flux in an iron and ROS-dependent manner. Flow cytometry analysis of autophagosomes (MDC staining) (a) and Western blotting analysis of p62 expression level (b) in ZnO NPs-induced PC12 cells for 6 h with or without TPEN pre-treatment for 30 min. Effects of 3-MA and SP600125 on intracellular zinc ions detected by zinc ions indicator Fluor™ Zn-520, AM staining (c) and ROS level detected by DCFH-DA staining (d) in ZnO NPs-treated PC12 cells. Cells were pretreated with 3-MA or SP600125 for 1 h, followed by exposure to 15 μg/mL ZnO NPs for 6 h. Data from at least three independent experiments were expressed as the means ± SD. ***p < 0.001 compared with the untreated control. e The inhibitory effects of NAC on the expression of p-JNK, LC3B conversion and p62 were determined by Western blotting. Cells were treated with 15 μg/mL ZnO NPs for 6 h in the presence or absence of NAC pre-treatment for 1 h. The densitometry of the above bolts from at least three independent experiments was shown in Figure S9C and D