Additional file 1 of Impaired anaplerosis is a major contributor to glycolysis inhibitor toxicity in glioma

Additional file 1: Supplemental Figure S1. POMHEX treatment results in accumulation of glycolytic metabolites upstream of enolase. Supplemental Figure S2. POMHEX treatment leads to an overall reduction in TCA cycle metabolites. Supplemental Figure S3. Pyruvate rescues POMHEX toxicity in glioma cells. Supplemental Figure S4. Rescue of POMHEX toxicity by anapleroticsubstrates. Supplemental Figure S5. Exogenously supplemented, supraphysiological levels of anapleroticsubstrates rescue POMHEX toxicity even in physiological PlasmaxTMmedium. Supplemental Figure S6. POMHEX treated cells are substrate limited for mitochondrial respiration which is attenuated by exogenous pyruvate. Supplemental Figure S7. Enolase inhibition induces bioenergetics stress preceding cell killing, which is rescued by exogenous pyruvate. Supplemental Figure S8. Exogenous pyruvate but not lactate rescues ATP production inhibited by enolase inhibitor treatment. Supplemental Figure S9. Glioma cells exhibit glutamine auxotrophyin vitro. Supplemental Figure S10. CB-839 toxicity is exaggerated under pyruvate free conditions and reversed by the addition of anapleroticsubstrates. Supplemental Figure S11. Metabolomic analysis of HEX treated ENO1deleted tumors from two different metabolomic platforms confirm elevation of glycolytic metabolites upstream, and reduction in the TCA cycle metabolites, downstream of the enolase reaction. Supplemental Figure S12. Exogenously supplemented pyruvate contributes to TCA cycle through both pyruvate carboxylase and pyruvate dehydrogenase reactions. Supplemental Figure S13. CO2levels modulate POMHEX toxicity. Supplemental Figure S14. Sensitivity of glioma cells to CB-839 is attenuated in physiological PlasmaxTMmedium. Supplemental Figure S15. Orally administered CB-839 is detectable in mouse plasma 2 hours post drug administration. via LC-MS (ESI).