Fig. 4: Glycolysis remains elevated with AOX-MR in COX-deficient T cells.

Splenic T cells were isolated and stimulated for 24 h as above. A Measurement of extracellular acidification rate (ECAR) in CD8⁺ and CD4⁺ T cells (2 × 10⁵ cells/well, N = 11–12/condition). Representative ECAR plots. B ECAR in Pan T cells after injection of oligomycin (filled) or LDHA inhibitor (50 mkM, empty). C Lactate in the medium of activated T cells (2 × 106 cells/ml) stimulated for 24 h (N = 3–5/condition). Lactate was measured by YSI analyzer in triplicate and averaged. Basal concentration was deducted from total production. D Hif1a staining by flow cytometry (N = 3–7/condition). E Schematic of stable isotope labeling with U-¹³glucose. Black circles indicate labeled carbons, white circles indicate unlabeled carbons. Created in BioRender. Mcguire, P. (2025) https://BioRender.com/zkz0xgp. F Quantification of pyruvate and lactate (m + 3) abundance deriving from glucose (N = 5–7/condition). G Quantification of succinate, fumarate, malate, and aspartate (m + 2) abundance deriving from glucose (N = 5–7/condition). H Quantification of citrate (m + 2) and citrate (m + 4) abundance deriving from glucose (N = 5 = 7/condition). Data are representative of two to three independent experiments and indicate mean and standard deviation. * p < 0.05, ** p < 0.01, *** p < 0.001 by one-way ANOVA and post-hoc Dunnett test against WT.