“…Protein synthesis was determined using our established methods. 28 , 34 Briefly, ∼30–50 mg of skeletal muscle tissue was homogenized 1:20 in isolation buffer (100 m m KCl, 40 m m Tris–HCl, 10 m m Tris Base, 5 m m MgCl 2 , 1 m m EDTA, 1 m m ATP, pH = 7.6) with phosphatase and protease inhibitors (HALT, Thermo Fisher Scientific) using a bead homogenizer (Next Advance Inc., Averill Park, NY, USA). After homogenization, subcellular fractions were isolated via differential centrifugation to obtain myofibrillar and collagen protein fractions.…”