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Isolation of mitochondria from tissues Freshly dissected brain tissue was cut into smaller pieces, rinsed in cold PBS, and homogenized using either a Dounce homogenizer or Waring blender in the presence of twice the tissue volume of buffer A (20 mM Hepes pH 7.4, 220 mM mannitol, 68 mM sucrose, 80 mM KCl, 0.5 mM EGTA, 2 mM Mg(Ac) 2 , 1 mM DTT, 1 protease inhibitor (Roche))