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200 l of MiR05 respiration buffer (110 mM D-sucrose, 60 mM lactobionic acid, 20 mM taurine, 20 mM Hepes, 10 mM KH2PO4, 3 mM MgCl2, 0.5 mM EGTA, and 1 g/l fatty acidfree BSA, pH 7.1, at 23C) (95, 96) was added to the pellet and gently mixed to make a mitochondrial suspension