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Western blot analysis Gastrocnemius homogenates were prepared in a non-denaturing buffer (2% Triton X-100, 300 mM NaCl, 20 mM Tris (pH 7.4), 2 mM EDTA, 2 mM EGTA, and 1% NP-40) containing the following phosphatase and protease inhibitors: 1 mM PMSF, 10 uM leupeptin, 38 uM aprotinin, 1 mM phenanthrolin, 1 uM pepstatin, 1 mM sodium fluoride, and 200 uM sodium vanadate