Fig. 6

The autophagy regulation role of JNK signaling pathway in ZnO NPs-treated cells. PC12 cells were pretreated with 10 μM SP600125 for 1 h prior to 6 h exposure to 15 μg/mL ZnO NPs. a The inhibitory effects of SP600125 on the expression of p-JNK and LC3B were determined by Western blotting. b Representative images and statistical analysis of PC12 cells with GFP-LC3 puncta. Cells were transiently transfected with GFP-LC3 plasmid. The inhibitory effect of SP600125 on autophagosome accumulation induced by ZnO NPs was imaged and calculated using the confocal microscopy (36 cells). Scale bar, 10 μm. c Time course analysis of phosphorylated and total Bcl-2 protein expression in ZnO NPs-treated PC12 cells. d Effects of JNK on ZnO NPs-induced phosphorylation of Bcl-2 and disruption of Bcl-2/Beclin 1 complex. Bcl-2 co-immunoprecipitation with Beclin 1 was detected with a monoclonal anti-Beclin 1 antibody. Input represented the total amount of p-Bcl-2 and p-JNK detected in whole-cell lysates by Western blotting analysis. Images were representative of at least three independent experiments, and the densitometry of the above bolts was shown in Figure S8E-G