Fig. 3: GSC display increased apoptotic priming and can be effectively killed by dual BCL-xL, MCL-1 inhibition.
From: Increased apoptotic sensitivity of glioblastoma enables therapeutic targeting by BH3-mimetics
A G7 and E2 GSC were treated with DMSO (ctrl), A-1331852 or ABT-737 as indicated for 16 or 24 h, respectively, harvested and protein expression was analysed by immunoblot. α-tubulin served as loading control. Representative image from n = 3 independent experiments shown. Quantification of n = 3 independent experiments, error bars represent mean ± SD (*p ≤ 0.0292) Welch’s test. B G7 or R24 GSC vectorCRISPR vs. MCL-1.1CRISPR and MCL-1.2CRISPR were treated with A-1331852 for 24 h and analysed for cell viability using an IncuCyte imager and SYTOX Green exclusion. Percentage cell death was calculated by normalising against maximal cell death verified by visual inspection. Error bars represent mean ± SEM from n = 3 independent experiments (*p ≤ 0.0449, **p ≤ 0.004, ***p ≤ 0.0006, ****p < 0.0001) Welch’s test. C E2, G1, G7, R24 GSC were treated with a combination of A-1331852 and S63845 in indicated concentrations for 24 h and analysed for cell viability using an IncuCyte imager and SYTOX Green exclusion. Percentage cell death was calculated by normalising against maximal cell death verified by visual inspection. Error bars represent mean ± SEM from n = 3 independent experiments (*p ≤ 0.0123, **p ≤ 0.0022, ***p ≤ 0.0004, ****p < 0.0001) Welch’s test. D Clonogenic survival assay of G7 GSC iRFP treated with indicated drugs 16 h after plating 250 cells per well. Colonies counted manually after 14 days. Error bars represent mean ± SD from n = 3 independent experiments (ns p = 0.1355, *p = 0.0243, **p = 0.0038) Welch’s test. Representative images of a replicate in one independent repeat scanned on LICOR imager. E E2 GSC vectorCRISPR and BAK/BAXCRISPR were treated as indicated for 2 h, harvested and protein expression was analysed by immunoblot. HSP60 served as loading control. Representative image from n = 3 independent experiments. F E2 and R15 GSC and paired DIFF cells were treated either with DMSO (grey) or a combination of A-1331852 and S63845 (both 0.1 μM) for 24 h and analysed for cell viability using an IncuCyte imager and SYTOX Green exclusion. Error bars represent mean ± SEM from n = 3 independent experiments. Representative IncuCyte images 24 h after treatment are shown.
