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. Author manuscript; available in PMC: 2015 Mar 28.
Published in final edited form as: Cancer Lett. 2013 Nov 19;344(2):272–281. doi: 10.1016/j.canlet.2013.11.005

Figure 5. mTOR inhibition-induced cell death was independent of caspase activation and blocked by autophagy induction.

Figure 5

(A) Apoptotic cell death induced by staurosporine was completely blocked by Z-VAD-FMK. MCC-2 cells were exposed to staurosporine (0.1μM) with or without Z-VAD-FMK (20 μM) for 6h followed by PI and Annexin V staining. (B) PP242-induced cell death was blocked by an autophagy inhibitor (bafilomycin-A1), but unaffected by a pan caspase inhibitor (Z-VAD-FMK). To confirm that PP242-induced cell death was caspase-independent, MCC-2 cells were treated with PP242 (2.5μM) and Z VAD-FMK (20μM) for 24 hours followed by PI and Annexin-V staining. To test if PP242-induced cell death could be blocked by autophagy, MCC-2 cells were treated with PP242 (2.5μM) for 18 hours and then with bafilomycin-A1(10nM) for additional 6 hours followed by PI and Annexin-V staining and flow cytometry analysis. Results shown are the means ± SEM of at least three independent experiments. *P < 0.05 compared with DMSO-treated cells. #P < 0.05 compared with staurosporine or PP242-treated cells.