(A) Schematic of the glycolysis model with chemical reactions and allosteric points of regulation described. Abbreviations: GLC—glucose, G6P—glucose-6-phosphate, F6P—fructose-6-phosphate, FBP—fructose-1,6,-bisphosphate, F26BP—fructose-2,6,-bisphosphate, GAP—glcyceraldehyde-3-phosphate, DHAP—dihydroxyacetone phosphate, BPG—1,3 bisphosphoglycerate, 3PG—3-phosphoglycerate, 2PG—2-phosphoglycerate, PEP—phosphoenolpyruvate, PYR—pyruvate, SER—Serine, GLY—glycine, Lac—lactate, MAL—malate, ASP—aspartate, Pi—inorganic phosphate, CI—creatine, PCI—phosphophocreatine, GTR—glucose transporter, HK—hexokinase, PGI—phosphoglucoisomerase, PFK—phosphofructokinase, ALD—aldolase, TPI—triosephosphoisomerase, GAPDH—glyceraldehyde-phosphate dehydrogenase, PGK—phosphoglycerate kinase, PGM—phosphoglycerate mutase, ENO—enolase, PK—pyruvate kinase, LDH—lactate dehydrogenase, MCT—monocarboxylate transporter, PDH—pyruvate dehydrogenase, CK—creatine kinase. (B) Overview of the algorithm and simulation method. (C) Measured values of the NADH/NAD+ ratio across a population of MCF10A breast epithelial cells. Three values of glucose concentration are considered (0.5 mM blue, 5.5 mM green, and 25 mM red). (D) Calculated fluxes (mM/hr) for glycolysis rate (Glycolysis) are defined as the rate of glucose to pyruvate (per molecule of pyruvate), pyruvate to lactate flux (LDH), rate of oxygen consumption (OxPhos), rate of NADH turnover (NADH), and ATP turnover (ATPase). (E) Calculated probability density function (PDF) of NAD+ concentrations. (F) Calculated probability density function (PDF) of NADH/NAD+ ratio. (G) Calculated probability density function (PDF) of ATP concentrations. (H) Calculated probability density function (PDF) of ATP/ADP ratio. (I) Box plots showing the distribution of concentrations computed from the simulation for each intermediate in glycolysis.