Fig. 3

SFN promoted autophagy in primary GMECs. A GMECs were treated with SFN (5 μmol/L) for the indicated times and the autophagosome was detected using the autophagy assay kit (Bar = 100 μm). B Fluorescence intensity analysis of autophagy shown in A, n = 30 randomly selected cells. One-way ANOVA, Dunnett's post-hoc test. C Autophagosome detection by transmission electron microscopy in GMECs treated with SFN (5 μmol/L) for 12 h. Arrows indicate the autophagosome. D–G Western blot analysis of LC3, Beclin1, Atg7 protein from 5 μmol/L SFN-treated GMECs for the indicated times (n = 3). One-way ANOVA, Dunnett's post-hoc test. H and I Western blot analysis of LC3 protein (n = 3). GMECs were pretreated with or without CQ (5 μmol/L) for 4 h in advance and then were incubated with SFN (1.25, 2.5 and 5 μmol/L) for 12 h. One-way ANOVA, Tukey's post-hoc test. J–O Western blot analysis of the autophagy-related signaling molecules from 5 μmol/L SFN-treated GMECs for the indicated times (n = 3). One-way ANOVA, Dunnett's post-hoc test. Data are presented as the mean ± SEM. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 and ^****P < 0.0001