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After treatment, the post-cultured medium was replaced with a 10% resazurin solution prepared in a DMEM-LG culture medium, and cells were incubated at 37 C for 2 h [37, 38]

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Start with MOTS-c alone at target dose for 1014 days, establish baseline response (measure fasting glucose, insulin sensitivity markers, and body composition changes), then introduce 5-Amino-1MQ at starting dose and titrate upward while monitoring