Extended Data Fig. 3: TKT deficiency does not alter Treg cellularity.

a, Flow cytometric analysis (left) and quantification (right) of Tregs from the thymus, spleen and pLN of 2-week-old WT and cKO mice. n=5 mice. b, Schematic diagram of chimeric Tregs in female Foxp3^Cre/+Tkt^fl/fl mice. c,d, Total cell number (c) and absolute CD4⁺ and CD8⁺ T cell number in the spleen and pLN (d) of 7-week-old chimeric WT or chimeric cKO mice. n=7 mice. e, Percentage of effector (CD44^hiCD62L^lo) cells in CD4⁺ T cells and CD8⁺ T cells in the spleen and pLN of 7-week-old chimeric WT or chimeric cKO mice. n=7 mice. f, Schematic representation of the bone marrow chimeric (BMC) experiment. g, Flow cytometric analysis of Tregs from the spleen and pLN of BMC WT and cKO mice. n=6 mice. h, Protein levels of TKT, Foxp3 and β-actin in CD4⁺CD8⁺ T cells and Tregs sorted from thymus and spleen of Foxp3^YFP-cre mice respectively (top). n=3 independent samples, Tregs is a positive control. mRNA expression of Tkt and Foxp3 during T cell development (bottom, data from the ImmGen RNA-seq database). T.DP.Th: double positive T cells in thymus; T.4.Th: CD4 single positive T cells in thymus; Treg.4.25.Sp: CD4⁺CD25⁺ T cells in spleen. i, Flow cytometric analysis (left) and quantification (right) of Tregs from 7-week-old Tkt^fl/fl and Cd4^CreTkt^fl/fl mice. n=5 mice. Data are representative of one (g) and at least two (a,c-e,h,i) independent experiments. Data are shown as mean ± s.d.. P value are determined by unpaired two-tailed Student’s t-test (c), and two-way ANOVA followed by Sidak’s multiple-comparisons test (a,d,e,g,i). ns, not significant. Numbers in gates indicate percentage of cells.

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