Fig. 3: BCL2 TMD mutants insert into the mitochondrial membrane.
From: Phenotyping of cancer-associated somatic mutations in the BCL2 transmembrane domain
A Schematic representation of the subcellular fractionation procedure to confirm protein membrane insertion. B Subcellular fractionation and carbonate extraction (Carb. extr) of mitochondria from HCT116 cells expressing BCL2 TMD mutants. Locations are controlled using Tom20 (mitochondrial fraction, M) and α-tubulin (cytosol, C). The mitochondrial fraction was treated with a sodium carbonate solution to remove loosely attached membrane proteins (soluble fraction, SP; membrane proteins, P). C Subcellular distribution of homo-oligomers of BCL2 TMD mutants in the mitochondria and ER. Representation of Pearson correlation coefficients between the BCL2 TMD constructs and Mitotracker or ER tracker. D Subcellular distribution of hetero-oligomers of BCL2 TMD mutants with BAX TMD in the mitochondria and ER. Representation of Pearson correlation coefficients between the BCL2 TMD constructs and Mitotracker or ER tracker. Data represented as mean ± SEM of at least n = 3. Significant differences compared to WT BCL2 TMD analyzed using Dunnett’s multiple comparison test (*p < 0.05, ****p < 0.0001).
