9.2.27PE and ABT-737 in Melanoma Figure 2. 9.2.27PE in combination with ABT-737 induces apoptosis in melanoma cells. (A) FEMX cells and Melmet-5 cells were treated with 9.2.27PE (100 ng/ml)6ABT-737 (10 mM) for 24 h. The combination treatment caused rounding of the cells and detachment from the surface of the flasks as compared to single agent treatment. (B) FEMX cells were treated with various doses of 9.2.27PE (1–100 ng/ml) or various concentrations of ABT-737 (0.1–10 mM) or a combination of 9.2.27PE (100 ng/ml) and ABT-737 (10 mM) for 24 h. Cells were subjected to Western Blot analysis for examination of the levels of PARP, Mcl-1, Caspase-3, BAX and a-tubulin. The blot is a representative of three individual experiments. (C) Melmet-5 cells were treated with 9.2.27PE (100 ng/ml)6ABT-737 (10 mM) for 24 h. Cells were subjected to Western Blot analysis for examination of the levels of PARP, Caspase-3, BAX and a-tubulin. To determine whether caspases and cathepsins were involved in the inactivation of PARP and activation of caspase-3, the cells were pre-treated for 1 h with Z-VAD-FMK (50 mM)6Z-FA-FMK (50 mM) before treatment with 9.2.27PE+ABT-737. The blot is a representative of three independent experiments. (D) Cell viability was measured in FEMX cells treated with 9.2.27PE (100 ng/ml)6ABT-737 (10 mM) for 24 h. To determine whether caspases and cathepsins were involved in the decreased cell viability caused by the combination of 9.2.27PE and ABT- 737, the cells were pre-treated with for 1 h with Z-VAD-FMK (50 mM)6Z-FA-FMK (50 mM). The data represents the mean 6 SD. doi:10.1371/journal.pone.0024012.g002 PLoS ONE | www.plosone.org 6 September 2011 | Volume 6 | Issue 9 | e24012