The TCA cycle mode switch toggles the fate of giant cells. (A) OCR in Mino-VEN-R parental and giant cells, normalized by protein amount. (B) Heat map of the TCA cycle–associated genes expression. A dichotomized profile of genes in the noncanonical (red) and canonical TCA cycle (blue). (C) Western blots of ACL in Mino-VEN-R cells treated with increasing doses of PBN. (D) Metabolites profiles of the TCA cycle intermediates. (E) Fraction enrichment of [U-¹³C]glucose–labeled intermediates (M+2) associated with TCA cycle was differentially regulated in P, G1, and R1 cells. (F) Heat map of representative nucleotides in P, G1, and R1 cells. (G) Coessentiality map of genes encoding metabolic enzymes, including 2 TCA cycle modes, clustered with purine metabolism. Correlation strength is indicated by the length and thickness of the connecting edges. (H) Fraction enrichment of [U-¹³C)]glucose–labeled pyrimidine intermediates (M+0 and M+5) was differentially regulated in P, G1, and R1 cells. (I) Expression of Got1 and Got2 was inversely regulated in parental Mino-VEN-R cells (P), PBN-treated (1 or 5 μM) cells, G1, and R1. Unpaired t test. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001. (J) Schematic illustration of the rewired TCA cycle in giant cells. ADP, adenosine diphosphate; AMP, adenosine monophosphate; CDP, cytidine diphosphate; CMP, cytidine monophosphate; CTP, cytidine triphosphate; dADP, deoxyadenosine diphosphate; dAMP, deoxyadenosine monophosphate; dATP, deoxyadenosine triphosphate; FCCP, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone; GDP, guanosine diphosphate; GMP, guanosine monophosphate; IMP, inosine monophosphate; ns, nonsignificant; UDP, uridine diphosphate; UMP, uridine monophosphate; UTP, uridine triphosphate; XMP, xanthosine monophosphate.