Differentiation of the cells to myocytes was induced by incubating the cells in a standard differentiation medium consisting of DMEM with 2% horse serum (Thermo Fisher Scientific) for 6 days, and was confirmed by upregulation of mRNA expression of Myod1 and Myog ( 2.4 Assessment of mitochondrial function and cell viability Mitochondrial function was assessed by measuring adenosine triphosphate (ATP) content and mitochondrial membrane potential using ATP Assay Kit-Luminescence and JC-1 MitoMP Detection Kit, respectively, following the manufacturers instructions (Dojindo Laboratories, Mashiki, Japan)
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