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7 Discussion In recent years, studies of immunometabolism have made it clear that amino acids are not merely substrates for protein synthesis, but are also embedded as multilevel metabolic nodes within the macrophage metabolic network, jointly shaping macrophage functional states through multiple layers, including substrate partitioning, TCA cycle anaplerosis, nutrient-sensing signals such as mechanistic target of rapamycin (mTOR) and GCN2, one-carbon/methyl donor cycling, and redox homeostasis ( At the level of the metabolic network, amino acids such as arginine, tryptophan, and glutamine can be regarded as key interfaces connecting multiple metabolic modules

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H.TraceyI.WesselyS.ArseneaultL.et al (2020)

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75 L of supernatants were transferred to a flat-bottom 96-well plate for measurement of free hemoglobin at OD 420 in a plate reader