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Tissues were homogenized and lysed in NETN buffer (20 mM Tris-HCl (pH 8.0), 100 mM NaCl, 1 mM EDTA and 0.5% Nonidet P-40) supplemented with 50 mM -glycerophosphate, 5 mM NaF and a protease inhibitor cocktail (Roche)