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Animal tissues were lysed with RIPA lysis buffer (50 mM Tris, 150 mM NaCl, 1% Triton X-100,1% sodium deoxycholate, 0.1% SDS, 5 mM sodium orthovanadate, 5 mM sodium fluoride, 1 mM and EDTA, pH 7.4) supplemented with 10 l/ml protease inhibitor cocktail (GE Healthcare Biosciences, Pittsburgh, PA, United States)