Fig. 4.

Effect of autophagy inhibitors on dioscin-induced human lung cancer cell death. A549 and H1299 cells were treated for 24 h with 5 μM dioscin with or without a 1 hour pretreatment of 5 mM 3-MA. The expression of LC3-II, cleaved caspase-3 and cleaved caspase-8 was detected by Western blotting (a), cell viability was determined by MTT assay (b), together with annexin-V and PI double-staining flow cytometry (c). Subsequently, A549 and H1299 cells were treated with different concentration of dioscin for 24 h with or without a pretreatment of 10 nM BafA1. Afterward, treated cells were subjected to MTT assay for cell viability (d), double-staining with annexin-V and PI and flow cytometry (e). Results are shown as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, dioscin versus 3-MA or BafA1 plus dioscin