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mnfr1738-sup-0001-figures1.tif291KFigure S1: Optimal amino acid concentrations facilitate leucine-induced myofibrillar but not total protein accretion. (A) C2C12 cells were differentiated for 5 days in 10% AA in DM or 30% AA in DM in the absence or presence of 5 nM IGF-I or 5 mM leucine. Medium and stimuli were refreshed every 48 h. Whole cell lysates were prepared. Subsequently, protein content was determined and protein levels were expressed as percentage of control. (B) C2C12 cells were differentiated for 4 days in the absence or presence of 5 mM leucine in 10% AA DM or 30% AA DM. Medium and leucine were refreshed every 48 h. Total RNA was extracted and cDNA synthesized. (B) MyHC-7 and (C) MyHC-4 gene expression levels were determined. (D) C2C12 cells were differentiated for 5 days in 10% AA in DM in the presence of 2 or 5 mM arginine. Whole cell lysates were prepared and MyHC-sl content was determined. * = Significant from respective control.

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