Fig. 5

Relationship of NEO212 induced autophagosome accumulation and apoptosis. a Cells were treated with 100 μM TMZ, POH, TMZ + POH, NEO212 or DMSO respectively for 48 h in A2780 and SK-O-V3 cells. Western blot analysis demonstrated BECN1, LC3B and ACTB expression; (b) Autophagic vacuoles in A2780 and SK-O-V3 cells treated with 100 μM NEO212 or DMSO were observed by transmission electron microscopy (TEM). The arrow indicates autophagic vacuoles. Scale bars: 0.5 μm. c The above drug-treated cells were inspected under confocal laser microscopy to detect LC3B puncta by immunofluorescence. Scale bars: 10 μm. Number of autophagic vacuoles was calculated using Fiji Image J software. d A2780 and SK-O-V3 cells treated with 100 μM NEO212 or DMSO were subjected to western blot to detected LC3B and ACTB expression in the presence or absence of Baf.A1. e-f A2780 and SK-O-V3 cells were treated with 100 μM NEO212 for 48 h with or without EBSS (e) or 3 M, Baf.A1 (f), and then apoptosis assay using Annexin-V &PI staining. The results shown are means ± SD; *p < 0.05; **p < 0.01; ***p < 0.001, NS = no significance