Fig. 6

Cholesterol-induced ROS initiate apoptosis and autophagy in TDSCs. a–f TDSCs were pretreated with NAC (5 mM, 1 h) before incubation with 10 mg/dL cholesterol and NAC for 24 h. The apoptosis proportion was evaluated by annexin V-FITC flow cytometry (a, b). Apoptosis-related proteins (cleaved caspase-3, Bax, Bcl-xL, and PARP) (c, d) and autophagy-related proteins (LC3-II and p62) (e, f) were analyzed by western blotting. g, h Cells were transfected with the mRFP-GFP-LC3 vector for 24 h and then pretreated with NAC (5 mM, 1 h) before incubation with 10 mg/dL cholesterol and NAC for 24 h. Representative images of fluorescent LC3 puncta are shown. Bar, 20 μm. All quantitative data are expressed as the means ± SEM of the results from three independent experiments. *p < 0.05 versus control, ^#p < 0.05 versus CHO. CHO, cholesterol