Fig. 3: MCL-1 is required for breast cancer cell line survival in vitro

a Western blot analysis of MCL-1 protein expression across a panel of human breast cancer cell lines. b, c MTS assay showing viability of MDA-MB-468 cells after 48 h treatment with indicated dose of MCL-1 inhibitor (b) UMI-77 or (c) A1210477. Bars indicate mean ± SD, of n = 3–5 independent experiments plated in triplicate. d Western blot showing full length (FL) and cleaved (cl) PARP following incubation of MDA-MB-468 cells with indicated doses of UMI-77 for 24 h. Actin as loading control shown below. e Western blot analysis (as for d) showing PARP cleavage and active caspase 3 (CC3) in MDA-MB-468 CRISPR/Cas9 edited for BAX/BAK deletion (see Supplementary Fig. S2B) or non-targeting control following 24 h treatment with 10 μM UMI-77 or 10 μM etoposide (etopo). Actin loading control is given for each membrane. f, g Incucyte Sytox Green cell death assay of cell lines described in e following 48 h treatment with 5 μM A1210477 (f) and 0.1 μM S63845 (g) in the presence or absence of 10 μM Q-VD-OPh caspase inhibitor. Cell death was calculated with the formula CD^treatment-CD^basal where CD^treatment is Sytox Green cells/cell confluence following 48 h treatment with MCL-1 inhibitor and CD^basal is Sytox Green cells/cell confluence in control samples at 48 h. Graph represents mean ± SEM from n = 3–4 independent experiments plated in triplicate