Venetoclax and S63845 synergize with sirolimus to induce apoptosis in human NF1/PTEN-deficient melanoma cells.a Western blots for NF1 and PTEN in a panel of human melanoma cell lines. HEK293 and Jurkart cells were included as positive and negative controls. The levels of total ERK1/2 expression serve as the loading control. b Relative cell viability of WM-3246 cells (Cell Titer Glo assay) upon treatment with sirolimus, venetoclax, or S63845 for 6 days. Mean ± s.d. values. c Relative cell viability of WM-3246 cells (Cell Titer Glo assay) upon treatment with the combination of sirolimus, venetoclax, and S63845 for 6 days. Mean ± s.d. values. d WM-3246 cell growth kinetics after treatment with the combination of sirolimus, venetoclax, and S63845 (for doses see panel c). Mean ± s.d. values. e Synergistic effects of venetoclax and S63845 on suppression of sirolimus-sensitized WM-3246 cells were analyzed by isobologram analysis. f Western blots for BCL2, BCLXL, and MCL1 in WM-3246 cells treated with venetoclax or S63845 for 24 h. g Western blots for cleaved caspase-3 in WM-3246 cells treated with the combination of sirolimus, venetoclax, and S63845.
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