(B) Abundance of total peptide (C13 and C12 combined) graphed after normalization to mean peptide abundance for all peptides mapping to glycolytic enzymes reveals consistent increase in expression of glycolytic enzymes under ischemia, despite overall reduction in translation. (C) Western blots demonstrating translational and post-translational alterations in the enzymes that metabolize pyruvate in the final step of glycolysis in cells surviving severe ischemia including an increase in the levels of LDHA and its activated, phosphorylated form. In addition, phosphorylation of pyruvate dehydrogenase to its inactive form was observed as a band shift toward a higher molecular weight. (D) Despite these alterations, metabolic analyses demonstrated a reduction in glucose uptake for cells surviving severe ischemia (p=.0004). (E) NMR spectroscopy of cell extracts incubated with U-13C-glucose demonstrated alterations in glycolytic flux consistent with the abovedescribed changes in protein expression with a significant reduction in the alanine:lactate ratio (p=.03). (F, G) Chemical inhibition of LDH (LDHi 737, 2.5 µM) resulted in the suppression of lactate excretion for HCC cells grown under standard and ischemic conditions within 24 hours of incubation; however, cells grown under ischemia were unable to survive these conditions when LDH was inhibited (*p