FIG. 6.

DIDS inhibits VDAC oligomerization and apoptosis. (A) HeLa cells were incubated with DIDS (100 or 200 μM) for 1 h, after which they were incubated with or without STS (1.25 μM; 2 h), harvested, cross-linked with EGS (250 μM; 15 min), and then analyzed by immunoblotting using anti-VDAC antibodies. The dramatic inhibition of STS-induced VDAC oligomerization by DIDS is shown. (B) Representative FACS analysis of apoptotic cell death, assayed using annexin V-FITC-propidium iodide staining. (C) Quantitative analysis of apoptosis, as measured by FACS analysis (from panel B). Results of one experiment representative of three similar experiments are shown. (D) Ratios of the BRET2 signals obtained for cells treated with STS (0.8 μM; 3.0 h) with or without pretreatment with DIDS (100 μM; 1 h). T-REx-293 cells expressing VDAC1 shRNA were cotransfected with plasmids encoding rVDAC1-Rluc (0.1 μg) and rVDAC1-GFP2 (0.8 μg). Data are represented as means ± standard errors of the means.