Extended Data Fig. 4: TKT depletion impairs Treg function by abolishing aTreg signature gene expression and transaldolase deficiency also dampens Treg function.
a, Schematic representation of the transfer colitis model for assessing Treg suppressive function in vivo by transferring WT or cKO Tregs into Rag1–/– recipients. b-e, In vivo Treg suppressive assay by a T cell-dependent adoptive transfer colitis experiment. b, n=5 mice; d, n=4 mice. (b), Representative ratio curve of post-transfer body weight relative to initial body weight of Rag1–/– recipients. (c), Representative H&E staining of distal colons of Rag1-/- recipients at 6 weeks after the transfer. Scale bars, 100 μm. (d), Representative flow cytometric plots of IFN-γ- or IL-17A-producing T cells in the spleen and colon (left). Right, cell number of IFN-γ- or IL-17A-producing T cells (gated from CD45.1+ TCRβ+CD4+ T cells). (e), Representative image of spleens from Rag1–/– recipients in (a). f, Schematic representation of the transfer colitis model for assessing Treg suppressive function in vivo by transferring BMC-WT and BMC-cKO Tregs into Rag1–/– recipients. g, RT-PCR analysis of mRNA levels of indicated genes in WT and Taldo1-deficient iTregs. n=3 cultures. h, Heatmap of the differentially regulated genes in chimeric WT Tregs relative to chimeric cKO Tregs isolated from 7-week-old female mice. Padj <0.05. i, Expression of CD44 (left, n=6 mice) and KLRG1 (right, n=5 mice) in chimeric WT and cKO Tregs from 7-week-old female mice (gated from CD4+CD25+YFP+ Tregs). j, Percentages of active Tregs (CD62LloCD44hi) in the spleen and pLN of 7-week-old chimeric WT or chimeric cKO mice (gated from CD4+CD25+YFP+ Tregs). n=6 mice. k, Genome browser views of indicated gene loci in chimeric WT and chimeric cKO Tregs. Data are representative of at least two independent experiments (b,d,g,i,j). ns, not significant. Data are shown as mean ± s.d. (b,g,i,j); box plots (d) 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), log-rank test (b) and two-way ANOVA followed by Sidak’s multiple-comparisons test (b,i,j). Numbers in gates indicate percentage of cells.
