Figure 2
Quantification of the apoptotic and the necrotic/late apoptotic population by annexin V/PI double staining and western blot following TMZ and fotemustine treatment. (A) Representative flow cytometry density plots of cells not exposed and exposed to 50 μ M TMZ or 32 μ M fotemustine after 144 h in the presence of O6BG (10 μ M) in the melanoma cell lines D14 and SK29, (B) ratio between apoptosis and necrosis/late apoptosis in TMZ- or fotemustine-treated cells after 144 h treatment in eight cell lines. For Malme 3M the TMZ time point was 196 h. (C, D) Western blot analysis of effector caspase-3 activation documented by the cleavage products p17 and p19, effector caspase-7 activation documented by cleavage product p20 and cleavage of PARP-1 protein (c-PARP-1) following treatment with 100 μ M TMZ and O6BG (10 μ M), or 32 μ M fotemustine and O6BG (10 μ M) in two representative melanoma cell lines D14 (C) and SK29 (D).
