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Mitochondrial preparations from C2C12 cells overexpressing FLAG-tagged full-length Alix were lysed with ice-cold lysis buffer (1 TBS [pH 8.0], 0.3% NP-40, 20% glycerol, 0.215 mM EDTA [pH 8.0], 150 mM KCl, protease inhibitors, and phosphatase inhibitors) by using a Dounce homogenizer (15 strokes)

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