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3T3-L1 pre-adipocytes cells were cultured with standard culture media (DMEM, 4.5 g/L glucose, L-glutamine, sodium pyruvate, 10% FBS, 1% antibiotic-antimycotic solution) and grown for 48 h before initiating differentiation using the manufacturer's suggested protocol and modified from previous published work

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Numerous studies have reported that NNMT has an inhibitory effect on the aerobic metabolism of sugars and fats, and NNMT Knockdown can increase oxygen consumption, promote sugar consumption and reduce fat synthesis, but the effects of NNMT on anaerobic energy metabolism have not been reported