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. 2017 May 18;18(5):1088. doi: 10.3390/ijms18051088

Figure 3.

Figure 3

The PI3K/AKT/mTOR pathway is involved in the regulation of both salinomycin-induced autophagy and apoptosis in prostate cancer cells. (A,B) AKT/mTOR expression. (A) PC-3 cells. (B) LNCaP cells. Cells were treated with 2 µM salinomycin for the indicated times. After treatment, total cell lysates were subjected to SDS-PAGE for Western blot analysis; (C) the effects of LY294002 on salinomycin-induced autophagy. Cells were pretreated with 10 µM LY294002 for 1 h, and acridine orange (AO) stained cells were evaluated by flow cytometry; (D) the effects of LY294002 on salinomycin-induced apoptosis. Cells were pretreated with 10 µM LY294002 for 1 h, and annexin-V-FITC stained cells were evaluated by flow cytometry; (E) the Effects of LY294002 on LC3, procaspase-3 and PARP protein expression. Cells were pretreated with 10 µM LY294002 for 1 h, and then the proteins were detected by Western blot analysis; and (F) the effect of LY294002 on caspase-3 activity. PC-3 cells were pretreated with 10 µM LY294002 for 1 h, and the proteins were detected by a caspase-3 colorimetric assay kit. All data were representative of at least three independent experiments. Data were shown as the mean ± SD. * p < 0.05.