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To ensure equal seeding density across wells, CFU validation was performed by plating aliquots on Middlebrook 7H11 agar and enumerating colonies after a 4-week incubation at 37 C

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In 3T3-L1 adipocyte cell models, 5-Amino-1MQ at 30 M concentration produced significant reductions in intracellular 1-MNA levels, confirming target engagement

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