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. 2014 Nov 22;9(3):675–688. doi: 10.1016/j.molonc.2014.11.005

Figure 6.

Figure 6

Combined treatment of nelfinavir and chloroquine selectively kills Tsc2−/− ELT3 cells. Tsc2−/− and Tsc2+/+ ELT3 cells were treated with either DMSO vehicle alone, 20 μM nelfinavir (NFV), 20 μM chloroquine (CQ), or both NFV and CQ and subjected to western blot and death assays. A) Following 3 h drug treatment, protein lysates were subjected to western blot analysis to determine levels of total and phosphorylated ‘(P)‐rpS6’, total and phosphorylated ‘(P)‐S6K1’, LC3‐II, SQSTM1, TSC2 and β‐actin. B) Quantification of cell viability. C) For quantification of cell death following 48 h of treatment, Tsc2+/+ and Tsc2−/− ELT3 cells were subjected to flow cytometry analysis following DRAQ7 staining. DRAQ7 exclusion (below line) represents the viable cell population, whilst positive DRAQ7 staining (above line) indicates cell death. The number of DRAQ7‐stained cells are graphed in D. E) Relative DNA fragmentation as determined by cell death ELISA. n = 3, *p = <0.05.