Fig. 6

APG-1387 induces autophagy in ovarian cancer cells. a The expression of LC3, Beclin1 and P62 was measured by western blot. Cells were treated with APG-1387(0, 3, 10, 30, 100, 300 nM) for 24 h. b Cells were transfected with GFP-LC3 plasmids, and then maintained in media with or without 3 nM APG-1387 for 24 h. The cells were then stained with DAPI and analyzed by fluorescence microscopy. c Statistical analysis of the percentage of LC3 puncta per cell. Columns, mean (n = 3); bars, SD. *P < 0.01 vs. untreated group. LC3 puncta per cell were quantified. d Cells were transfected with Beclin1 siRNAs. Western blot was used to detect the expression of Beclin1. e Cells were transfected with ATG7 siRNAs. Western blot was used to detect the expression of ATG7. f Cells were transfected with Beclin1 siRNAs. After 24 h treatment with or without 3 nM APG-1387, western blot analysis was performed for indicated proteins. g Cells were transfected with ATG7 siRNAs. After 24 h treatment with or without 3 nM APG-1387, western blot analysis was performed for indicated proteins. h Western blot analysis was performed for indicated proteins in cells transfected with siBeclin-1 and treated with 10 nM APG-1387. i Western blot analysis was performed for indicated proteins in cells transfected with siATG7–1 and treated with 10 nM APG-1387