Metabolic analysis of thymi from PTENΔ/Δ, ACLYΔ/Δ, PTENΔ/Δ;ACLYΔ/Δ or PTENflox2;ACLYflox2 mice. (A) Principal component analysis of 4 PTENΔ/Δ mutant, 3 ACLYΔ/Δ mutant, 3 PTENΔ/Δ;ACLYΔ/Δ, and 5 Ptenflox2;Aclyflox2 thymi from preleukemic 8-week-old mice undergoing unsupervised metabolic analysis. (B-D) Heat maps of the 50 most deregulated metabolites (B), analysis of glycolytic intermediates, Krebs cycle intermediates, glutamine, glutamate, citrate, CoA, acetyl-CoA and fatty acids (C), and pathway analysis in 4 PTENΔ/Δ mutant, 3 ACLYΔ/Δ mutant, 3 PTENΔ/Δ;ACLYΔ/Δ, and Ptenflox2;Aclyflox2 thymi (D). (E-F) Extracellular acidification rate (ECAR) (E) and oxygen consumption rate (OCR) (F) of sorted DP thymocytes in 2 independent experiments, with at least 2 per group. Adj, adjusted; aKG, α-ketoglutarate; F1,6DP, fructose 1,6 diphosphate; FAS, fatty acid synthesis; FCCP, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone; FDR, false discovery rate; TCA, tricarboxylic acid cycle; WT, wild-type.