FIGURE 3.
ICER regulates glycolysis in Tregs. (A) Sorted Tregs from SLOs of Foxp3 cre and ICER fl/fl Foxp3 cre mice were stained with CTV, expanded for 96 h, and stained for flow cytometry analysis. (B) Sorted naive T cells from ICER fl/fl and ICER fl/fl Foxp3 cre mice were expanded in iTreg‐polarising conditions for 96 h and extracellular acidification rate (ECAR) was assessed by Seahorse analysis. (C) Quantification of basal glycolysis, glycolytic capacity, and glycolytic reserve. (D) OCR/ECAR ratio was determined from basal respiration and glycolysis rates. (E) Western blot analysis of mTOR phosphorylation in WT and ICER‐deficient iTregs after 96 h of differentiation (left). Quantification (right). (F) Sorted naive T cells from ICER fl/fl and ICER fl/fl Foxp3 cre mice were expanded in iTreg‐polarising conditions for 96 h, 100 nM Rapamycin was added after 72 h, and the extracellular acidification rate (ECAR) was assessed by Seahorse analysis. (G) Quantification of basal glycolysis, glycolytic capacity, and glycolytic reserve. Student's t‐test (A, D) and Two‐way ANOVA with multiple comparisons (C, G) were used for statistical analysis. Dots represent biological replicates. Mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, ns, not significant.