The flux analysis used metabolite data collected between days 12 and 14 post-induction, because the newly differentiated adipocyte were expected to exhibit the mature phenotype by this time. Figure 5 summarizes the estimated fluxes through the major pathways. The complete flux data are shown in Table S2. Overall, there were 18 reactions significantly affected by both UCP1 expression and FCCP treatment. These reactions participated in the following pathways: glycolysis, PPP, lipid metabolism and amino acid metabolism. UCP1 expression and FCCP treatment increased glucose uptake by 85 and 95%, respectively. Fluxes through reactions in glycolysis increased as well. The most substantial flux increase involved lactate dehydrogenase. UCP1 expression and FCCP treatment increased lactate output respectively. Mitochondrial Complanatoside-A Uncoupling had the opposite effect on the PPP. UCP1 expression and FCCP treatment significantly decreased the net flux through PPP by 81 and 100%, respectively. The flux estimates also pointed to a significant down-regulation of lipogenesis. Phosphoenolpyruvate carboxykinase catalyzes the conversion of cytosolic oxaloacetate into phosphoenolpyruvate, a key step in glycerogenesis generating the glycerol moiety of TG. Uncoupling protein-1 expression and FCCP treatment decreased the flux through PEPCK by 36 and 78%, respectively. Uncoupling also reduced the supply of cytosolic acetyl-CoA units for de novo fatty acid CH5138303 synthesis. UCP1 expression and FCCP treatment lowered the transport of citrate from the mitochondria into cytosol and its subsequent cleavage into acetyl-CoA and OAA by 42 and 78%, respectively. Overall, UCP1 expression and FCCP treatment depressed the rates of TG formation, accumulation and lipolysis by 40,52% and 15,80%, respectively. The changes in amino acid metabolism were relatively small in magnitude. The largest effects were on the production of proline and alanine. UCP1 expression and FCCP treatment elevated alanine production by 70 and 249%, respectively. Proline formation was nearly zero in the unmodified and untreated control cells.