Ab180715

Antibodies used in the experiments were anti-CypD (1:1000, ab110324, Abcam, Cambridge, MA, United States); anti-VDAC1 (1:1000, ab154856, Abcam); anti-ANT (1:1000, ab180715, Abcam); and anti-β-Actin (1:10,000, Sigma, St. Louis, MO, United States). See Supplementary Material for details.

Western blotting procedures were completed as previously reported [6 (link), 8 (link), 36 (link)] using rodent OXPHOS Cocktail, ab110411, Abcam, Cambridge, UK, 1:250 dilution), VDAC2 (Santa-Cruz, 32059, Dallas, TX, 1:1000 dilution) and adenine nucleotide translocase 1 (ANT1, ab180715, Abcam, 1:1000 dilution) antibodies. Briefly, protein content was determined using BCA protein assay kit (Life Technologies, Carlsbad, CA, USA) to prepare samples. Proteins were separated on a 12% acrylamide gel and then transferred onto a low-fluorescence polyvinylidene difluoride membrane that was then blocked and incubated with the respective primary antibodies listed above overnight at 4°C. Membranes were then washed and incubated with their corresponding infrared fluorescent secondary antibodies (LI-COR, Lincoln, NE, USA) and imaged with the LI-COR infrared imager. Membranes were analysed with ImageJ software. Proteins were made relative to total protein measured on a separate membrane stained with amido black stain where loading accuracy was tested with a coefficient of variation of 8.6%.