Figure S2.

Loss of Arkadia in T cells results in elevated thymic Treg cells but no perturbations in the intestinal T cell compartment of SPF mice, related to Fig. 2. (A) Analysis of thymocyte subsets from control littermates and Arkadia conditional KO mice. Representative flow cytometry panels (left), and proportions and total numbers of double-positive thymocytes from multiple mice (right). (B) Analysis of nTreg cell proportions and numbers among CD4 single-positive T cells from different lymphoid organs of control littermates and Arkadia conditional KO mice. (C) Analysis of GATA3 and FOXP3 expression in CD4⁺ TCRβ⁺ T lymphocytes from LILP of control and Arkadia conditional KO mice under SPF conditions. (D) INF-γ and IL-17A production by CD4⁺ TCRβ⁺ T lymphocytes from LILP of control and Arkadia conditional KO mice in SPF conditions. Data for C and D are from one of three independent experiments, with n = 13 in the three experiments. Unpaired t test was used for statistical analysis. Black circle, control littermates; red triangle, Arkadia conditional KO mice (A–D). (E) Mice were the same as those in Fig. 2 C. CD4⁺ CD8⁻ T lymphocytes from different tissues were stained for Helios and FOXP3 to analyze the development of nTreg cells. Relative ratios of frequency and cell number of nTreg cells (Helios⁺ FOXP3⁺) are shown, respectively. The relative ratios of cell number were normalized to the ratios of donor B cells in peripheral blood. C1 LN, cecal-colonic draining LN; control/WT, control cell frequency or cell number divided by that of WT cell (black circle); Ark KO/WT, Arkadia mutant cell frequency or cell number divided by that of WT cell (red triangle). Data were obtained from a single experiment with a total of 13 mice for two experimental groups. Ratios of cotransferred cells in each animal were calculated individually and combined for analysis with unpaired t test. All error bars represent SD (A–E).  *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.