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Samples were diluted with 10 volumes of ice-cold EB (100 mM KCl, 2.5 mM MgCl 2 and 50 mM HEPES-KOH pH 7.5) containing 0.25% NP-40 and centrifuged through a 0.5 M sucrose layer at 10,000 g for 5 min at 4 C