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Real-time quantitative PCR (RT-qPCR) The total RNA was extracted from leukocytes using the QIAamp RNA Blood Kit (Qiagen, Hilden, Germany), and cDNA was produced with the High-Capacity cDNA Reverse Transcription Kit (Thermo Fisher, Waltham, MA) as previously described 18

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Specifically, adding 5-amino-1mq (30 M and above) to C2C12 myoblast cultures (a common in vitro myoblast model) markedly upregulates the expression of myogenic marker genes, including myosin heavy chain (MyHC), actin, and myogenin (MyoG)

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