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. 2023 Nov 28;11:1293268. doi: 10.3389/fbioe.2023.1293268

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Copyright © 2023 Hu, Wang, Bryant, Liu, Xie, West and Walsh.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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The roles of NADH and FAD in metabolic pathways. NADH and FAD are coenzymes in glycolysis, the TCA cycle, glutaminolysis, and the electron transport chain. 2-DG inhibits glycolysis, forcing a cell to use alternative pathways for metabolism. Cyanide inhibits complex IV of the electron transport chain, effectively inhibiting OXPHOS. BPTES inhibits glutaminolysis. Each metabolic pathway varies the fluorescence lifetimes of NAD(P)H and FAD differently. IRR = intensity redox ratio FAD/(FAD + NAD(P)H), FLIRR = fluorescence lifetime redox ratio NAD(P)H α ₂/FAD α ₁.