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Liver proteins were extracted by RIPA lysis buffer and western blot was performed as previously described.²⁶ The primary antibodies used were the following: rabbit anti‐PGC‐1α (1:1000 dilution, wl02123, Wanlei); rabbit anti‐NRF1 (1:5000 dilution, ab175932, Abcam); rabbit anti‐TFAM (1:1000 dilution, ab252432, Abcam); rabbit anti‐MFN‐1 (1:1000 dilution, abs120025, absin); rabbit anti‐MFN‐2 (1:5000 dilution, ab124773, Abcam); rabbit anti‐OPA‐1 (1:3000 dilution, ab157457, Abcam); mouse anti‐DRP‐1 (1:500 dilution, sc‐271583, Santa Cruz); mouse anti‐Fis‐1 (1:500 dilution, sc‐376447, Santa Cruz); mouse anti‐Bax (1:500 dilution, sc‐23959, Santa Cruz); rabbit‐anti‐Bcl‐2 (1:500 dilution, wl01556, Wanlei) and rabbit anti‐β‐actin (1:5000 dilution, D110001, Sangon Biotech). The nitrocellulose membrane was incubated overnight at 4°C with the above primary antibodies and then incubated with the appropriate horseradish peroxidase (HRP)‐conjugated secondary antibodies at room temperature for 2 h. The bands were visualized using Tannon 5200 (Tanon Science & Technology Co., Ltd.) and quantified by Image J software (National Institutes of Health).

Exposure of the 6A7 epitope of BAX was assessed by immunoprecipitation with a 6A7-domain-specific antibody purchased from Santa Cruz (SC-23959). Protein G beads (50 µL, Santa Cruz) were washed three times with 3% BSA in PBS and incubated with 15 µL 6A7 antibody at 4 °C for 1 hr. Recombinant full-length BAX (10 µM) was incubated with four equivalents of BIM–BH3 peptide alone and in the presence of five or ten equivalents of EO for 15 min at room temperature. Incubation of full-length recombinant BAX with 0.1% Triton X served as a positive control for exposure of the 6A7 epitope. After incubation, 10 µL of each reaction was transferred to the Protein G beads pre-loaded with anti-6A7 antibody and 1 µL was reserved as a loading control. After 90 min of incubation at 4 °C, beads were collected and washed three times with 500 µL of 3% BSA in PBS and solubilized with 25 µL LDS/DTT loading buffer. Samples were resolved by SDS-PAGE electrophoresis and western blot analysis with a BAX monoclonal antibody (Cell Signaling, Cat. # 2772) 1:1000.