Ab42364

All chemicals were of the highest grade of purity commercially available and purchased from Sigma, UK unless stated otherwise. Aqueous solutions were prepared in ultrapure water, while for non-aqueous solutions, ethanol or DMSO was used as solvent instead. Antibodies were all commercially available and used following suppliers’ recommended protocol or validation profile for the given application unless otherwise stated. Antibodies used in Western blotting (antibody, supplier, catalogue number, dilution): DRP1, BD Biosciences, 6111113, 1:1000; phospho-DRP1 S616, CST, 4494, 1:1000; β-actin, Sigma, A2228, 1:10000; OPA1, Abcam, ab42364, 1:500; MFF, Proteintech, 17090-1-AP, 1:500; FIS1, Proteintech, 10956-1-AP, 1:1000; GFP, Sigma, G1544, 1:2000. Antibodies used in immunocytochemistry (antibody, supplier, catalogue number, dilution): Tubulin-α, BioRad, MCA78G, 1:400; β-actin, Sigma, A2228, 1:1000

Cells with indicated treatments were lysed in IP buffer supplemented with protease inhibitors cocktail (B14001, bimake). Then lysates were incubated with pre-linked anti-OPA1 antibody (Abcam, ab42364), HSP90 (13171-1-AP, Proteintech), or control rabbit IgG (Abcam, ab172730) Dynabeads protein G (Life technologies, 10004D) for 2 h at room temperature, following immunoblot with indicated antibodies.