Fig. 5: Bak is required for VDAC2-dependent tBid sensitivity.
A Representative time course measurements of ΔΨm in permeabilized mouse hepatocytes infected with adenovirus encoding VDAC2 or GFP and treated with 3.7 nM tBid (left) and dot plot showing the changes in TMRE fluorescence during the treatment period (n = 3 experiments with 1 or 2 technical replicates per condition; p = 0.049, two-tailed t-test). Left inset shows the cyto c release by western blot of the cytosolic fraction. B As in (A) but using hepatocytes of a Bak-/- mouse. High digitonin (600 µg/ml) was used as a reference for cyto c release in the western blot. Dot plot: n = 2 experiments with 1 or 2 technical replicates per condition. C Representative immunoblots of VDAC2 and Bak in the membrane fractions of permeabilized WT and Bak-/- hepatocytes from (A) and (B); Tim23 as loading control; VDAC2 blots are cropped between the 25 and 37 kDa molecular weight markers D Representative time-course ΔΨm measurements in control and Bak shRNA treated HepG2 cells treated with 0.5 nM tBid after removal of the cytosol. Traces are normalized between the initial and fully depolarized (FCCP) values. E Immunoblots of cyto c in the supernatant of the permeabilized control and Bak shRNA treated cells; * indicates an empty lane. (D–E) The experiments were repeated 4 times with similar results. Source data are provided in the Source Data file.
