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Immunoblot analysis Whole cell lysates were prepared using radio-immunoprecipitation assay (RIPA) lysis buffer (pH 7.8) containing 25 mM Tris-HCl, 120 mM NaCl, 5 mM EDTA and 0.5% NP-40 supplemented with 100 μg/mL phenylcmethylsulfonyl fluoride, 200 mM sodium orthovanadate, 0.5 M NaF, protease inhibitor cocktail sets III and IV and phosphatase inhibitor cocktails 2 and 3 (Sigma-Aldrich)