Fig. 4: Single amino acid mutations in the BCL2 TMD impair BCL2 apoptotic function.
From: Phenotyping of cancer-associated somatic mutations in the BCL2 transmembrane domain
A The anti-apoptotic activity of BCL2 TMD mutants in response to BAX TMD-induced cell death measured by caspase 3/7 activity in HCT116 cells co-transfected with VN BCL TMD constructs. Data represented as mean ± SEM of at least n = 6. Significant differences compared to BCL2 TMD WT analyzed using Šídák’s multiple comparison tests (**p < 0.01, ***p < 0.001). NT = non-transfected; MOCK = cells transfected with empty VN plasmid. B Quantification of HCT116 cell viability measured by crystal violet staining. Data represented as mean ± SEM, n = 4. Significant differences compared to the BCL2 TMD WT analyzed using Šídák’s multiple comparison tests (****p < 0.0001). C The anti-apoptotic activity of BCL2 FL mutants in response to BAX TMD-induced cell death measured by caspase 3/7 activity in HCT116 cells co-transfected with BCL2 FL mutants. Data represented as mean ± SEM of at least n = 3. Significant differences compared to the BCL2 FL WT analyzed using Šídák’s multiple comparison tests (**p < 0.01, ***p < 0.001). NT = non-transfected; MOCK = cells transfected with pCDNA3.1 empty plasmid. D Quantification of HCT116 cell viability measured by crystal violet staining. Data represented as mean ± SEM, n = 4. Significant differences compared to the BCL2 TMD WT analyzed using Šídák’s multiple comparison tests (**p < 0.01, ****p < 0.0001). E The anti-apoptotic activity of BCL2 FL mutants in response to BAX FL-induced cell death measured by caspase 3/7 activity in HCT116 cells co-transfected with BCL2 FL mutants. Data represented as mean ± SEM of at least n = 3. Significant differences compared to the BCL2 FL WT analyzed using Šídák’s multiple comparison tests (**p < 0.01, ***p < 0.001). NT = non-transfected; MOCK = cells transfected with pCDNA3.1 empty plasmid. F Caspase 3/7 activity was measured in HCT116 cells transfected with BCL2 FL WT or I230V mutant and treated with 1 µM staurosporine for 24 h. Data represented as mean ± SEM (n = 3; *p < 0.05, **p < 0.01, ***p < 0.001). NT = non-transfected/non-treated, MOCK = cells transfected with pCDNA3.1 empty plasmid.
