Fig. 3: Hypertonicity increases transcription of BCL-2 family proteins without affecting interaction of BCL-XL/MCL-1 with BAX.

a HCT116 cells were challenged with NaCl (60 mM) for the indicated periods. mRNA levels of genes encoding the indicated proteins were analyzed by qPCR. Shown are data points and mean from two independent experiments. b, c HCT116 cells were challenged with NaCl (60 mM), WEHI-539 (1.25 µM) and S63845 (2.5 µM) for 5 h. After washing and cell lysis, immunoprecipitation was performed with antibodies specific for b BCL-XL and c MCL-1. Immunoprecipitates were analyzed together with the corresponding lysates by western blotting using antibodies specific for the indicated proteins. The BCL-XL inhibitor WEHI-539 and the MCL-1 inhibitor S63845 served as positive/negative controls. d HCT116 cells were challenged with NaCl (60 mM) for 24 h or left untreated. Immunoprecipitation was performed as described in (b). For (b–d), data shown are representative of at least two experiments performed. e HCT116 cells and BID-, BIM-, PUMA-, BAX-, and BAK-deficient variants thereof were challenged with the indicated concentrations of WEHI-539 for 18 h in the presence and absence of NaCl (60 mM). Data points and mean ± SEM from three independent experiments are shown.