Fig. 6

ASP diminished chondrocyte apoptosis via activation of autophagy. Chondrocytes were pretreated with 3-MA (10 μM), CQ (20 μM), and SCH772984 (10 nM) for 4 h before ASP treatment followed by SNP (1 mg/ml) for 24 h. a Chondrocyte apoptosis was measured by flow cytometry. b The formation of autophagosome was detected by LC3II immunostaining. c Levels of LC3II/I and p62 in chondrocytes after stimulation were measured by western blot. d Quantitative analysis of LC3II in chondrocytes. e Quantitative analysis of p62 in chondrocytes. f Representative microscopy images of tfLC3 puncta in chondrocytes stained for autophagosomes (green) and autolysosomes (red) as an indicator of autophagy flux. Autophagy increased in the ASP-treated groups in a dose-dependent manner. The data are presented as the mean ± SD of three independent experiments (n = 3). ***p < 0.001, ^#p < 0.0001, and statistical significance was determined by one-way ANOVA