Fig. 6: Selective killing of tumor cells by pharmacological targeting of VDAC2-Bak dependent cell death. | Nature Communications

Fig. 6: Selective killing of tumor cells by pharmacological targeting of VDAC2-Bak dependent cell death.

From: VDAC2 and Bak scarcity in liver mitochondria enables targeting hepatocarcinoma while sparing hepatocytes

Fig. 6

A Schematic representation of the pharmacological targeting strategy-employing an inhibitor of anti-apoptotic Bcl-2 family protein Mcl-1, S63845 together with an activator of the Bid pathway (TRAIL), tumor cells can be selectively killed. B Western blots of cyto c release in intact HepG2 and primary hepatocytes treated with 300 nM S63845 for 48 h with TRAIL (50 or 200 ng/ml) added in the final 3 h. The fraction of release, relative to high digitonin, was estimated by densitometry analysis of the western blot bands (% values above each band). Similar results were found in two biological replicates. C TRAIL dose-response curves of cell viability for HepG2 and hepatocytes with and without 48 h pre-treatment with 300 nM S63845. Cells were treated with TRAIL at the indicated concentrations for 8 h and viability was assessed by imaging propidium iodide exclusion. N = 5 biological replicates (3 technical replicates each) for 0 and 50 ng/ml TRAIL and 4 biological replicates (3 technical each) for 100 and 200 ng/ml TRAIL; statistical significance calculated by two-way ANOVA with Holm-Sidak post-hoc pairwise comparisons; statistically significant interaction between S63845 and TRAIL: p < 0.001. See p-values for pairwise comparisons in Supplementary Data 1. D Time dependence of cell killing by TRAIL (50 ng/ml) in cells with or without S63845 pre-treatment (300 nM, 48hrs). N = 2, 3 and 5 biological replicates at 2 h, 4 h, and 8 h, respectively, with 3 technical replicates within each experiment; statistical significance for the effects of time and S63845 was calculated by two-way ANOVA with Holm-Sidak post-hoc pairwise comparisons; statistically significant interaction between S63845 and time with 50 ng/ml TRAIL: p < 0.001. See p-values for pairwise comparisons in Supplementary Data 1. E TRAIL dose-response curves of WT and VDAC2KO HepG2 cell viability after 48 h pre-treatment with varying concentrations of S63845. Cells were treated with TRAIL at indicated concentrations for 4 h and viability was determined by luminescence assay, normalized to solvent-treated cells (N = 2 biological replicates). F TRAIL dose-response curves of WT and VDAC2KO PLC cell viability after 48 h pre-treatment with varying concentrations of S63845. Cells were treated with TRAIL at indicated concentrations for 4 h and viability was determined by luminescence assay, normalized to solvent-treated cells (N = 3 biological replicates). C–F Data are plotted as means ± SEM of multiple experiments; *p < 0.05, **p < 0.01, ***p < 0.001, ns: not significant; details of statistical analyzes and p-values are compiled in Supplementary Data 1. Source data are provided in the Source Data file.

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