Fig. 6: Metabolomic adaptations of naïve Ant2^-/- CD8⁺ T cells.

A–O Comparison of area under the curve (AUC) values from metabolomics analysis using ¹³C₆-glucose (red) or ¹³C₅-glutamine (blue) tracing alongside Label-Free Quantitation (LFQ) values from proteomics analysis in naïve (CD44⁻) WT and Ant2^−/− CD8⁺ T cells. A Bar graph showing the ratio between the AUC values of unlabeled Phosphocreatine (Pcr) and Creatinine (n = 6). B Bar graph displaying the LFQ values of Creatine Kinase (WT: n = 4, Ant2^−/−: n = 5). C Bar graph demonstrating the NAD⁺/NADH ratio (n = 6). D–H LC-MS measurements of labeling patterns for glycolytic derivatives after [¹³C₆]-glucose tracing. Bar graphs display AUC values of alanine m + 3 (D, F), and pyruvate m + 3 (E, G) in the cell and media fractions, respectively, and lactate m + 3 in the media fraction (n = 6) (H). Bar graphs showing LFQ values of P5CS (I) and PYCR2 (J) (WT: n = 4, Ant2^−/−: n = 5). K–N Bar graphs displaying AUC from LC-MS measurements of proline labeling patterns (n = 6). K Bar graph showing AUC values for ¹³C_6-glucose-derived proline (m + 2) in the cell fraction. L, M Bar graphs showing AUC values for ¹³C₅-glutamine-derived proline (m + 3 and m + 5) in the cell and media fraction, respectively, and AUC values for proline (m + 5) in the media fraction (N). O Heatmap illustrating differentially expressed unlabeled nucleotides and their precursors. The heatmap was generated based on the log2 fold change of individual metabolites (n = 5). P Bar graphs illustrating the (m + 3) to (m + 2) ratio of specific TCA-cycle intermediates and TCA-cycle-derived metabolites in naïve (CD44⁻) WT and Ant2^−/− CD8⁺ T cells (n = 6). Q Representative immunoblot analysis of whole-protein extracts from purified WT and Ant2^−/− CD8⁺ cells using anti-tPDH and pPDHser300 antibodies. Anti-tubulin was used as a loading control (pooling of n = 3). Statistical method: two-tailed Mann–Whitney test (A–N, P), or Student’s t test analysis conducted on log₂-transformed data after the Z-score normalization step (O). Data represented as the mean ± S.D. n refers to the number of biologically independent samples. ns = not significant. Source data are provided as a Source Data file.