Extended Data Fig. 5: Loss of TKT leads to metabolic reprogramming, defective mitochondrial function and impaired energy production.
a, Analysis of ECAR of WT and cKO Tconvs isolated from Tktfl/fl and Cd4CreTktfl/fl mice. n=4 cultures. b, Analysis of ECAR of WT and cKO iTregs. n=5 cultures. c, Analysis of OCR of WT and cKO iTregs. WT iTregs: n=6 cultures; cKO iTregs: n=5 cultures. d, Relative isotopomer abandance of M+2 citrate, α-KG, succinate, fumarate and malate in WT and cKO Tregs following incubation with [13C6]glucose for 12 hours. n=3 independent samples. e, LC-MS/MS analysis of amino acids in the serum of 3-week-old WT and cKO mice. FC means fold change. n=8 mice. f, Representative quantifications of MitoSOX, MitoTrackerTM Red CMXRos and Mitotracker Green in WT and TKT-deficient Tregs by flow cytometry. n=5 mice. Tregs for MitoSOX and MitoTrackerTM Red staining were isolated from 3-week-old WT and cKO mice. Tregs for Mitotracker Green staining were isolated from 7-week-old Tktfl/fl and Cd4CreTktfl/fl mice. g, Level of ATP in WT and cKO Tregs isolated from 2-3-week-old mice. n=6 independent samples. h, Representative electron micrographs of mitochondria and quantification of mitochondrial area and cristae width of WT and cKO iTregs induced in vitro. Scale bars, 1 μm and 200 nm, respectively. n=10 independent samples. i, Western blotting analysis of levels of indicated proteins in WT and TKT-deficient Tregs. j, Representative ratio curve of post-transfer body weight relative to initial body weight of Rag1–/– recipients in the adoptive transfer colitis experiment supplied with or without NAC in drinking water. Water: WT Tregs: n=6 mice, cKO Tregs: n=7 mice, No Tregs: n=5 mice; NAC: WT Tregs: n=6 mice, cKO Tregs: n=7 mice, No Tregs: n=8 mice. P value were analyzed by using the body weight changes at 7 week. k, Representative IFN-γ production in CD45.1+CD4+ T cells of Rag1–/– recipients in the adoptive transfer colitis experiment supplied with or without NAC in drinking water. n=5 mice. Data are representative of three (a,b,c,h,i) and one (d,e,f,g) independent experiments. Data are shown as mean ± s.d. (a-d,f-h,k); box plots (e) include minima, maxima, center line, median; box limits, upper and lower quartiles; whiskers, 1.5x interquartile range; points, outliers. P value are determined by unpaired two-tailed Student’s t-test (g,h,j), and two-way ANOVA followed by Sidak’s multiple-comparisons test (a-d,f,k).
