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The cellular pellet was resuspended in cell lysis buffer containing 20.4 mM ethylenediaminetetraacetic acid (EDTA), 146.9 mM ethylene glycol-bis (2-aminoethylether) N, N,N, N-tetraacetic acid (EGTA), 1 mM calcium glycerophosphate, 1 mM sodium orthovanadate, 2.6 mM sodium pyrophosphate, 20.4 mM Tris-HCl and 1% (v/v) Triton X-100, supplemented with 0.1 mM phenylmethanesulfonylfluoride (PMSF), 12% (v/v) protease inhibitor cocktail (Sigma #P8340), 1 mM dithiothreitol (DTT) and 20 mM sodium fluoride

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Together, they raise downstream IGF-1, which is one of the actual drivers of muscle protein synthesis and tissue repair