Deletion of IL-4Rα on Foxp3⁺ Treg cells does not break immune tolerance under a steady state.

(A) Representative flow cytometry analysis of CD4+ Foxp3+ cells among CD3+ cells from spleen, lung, MLN, and thymus of naïve IL-4Rα−/lox and Foxp3cre IL-4Rα−/lox mice. (B) Frequency of CD4+ Foxp3+ T cells from (A). (C) Body weight of naïve IL-4Rα−/lox and Foxp3cre IL-4Rα−/lox mice. (D) Organ weights of naïve mice. (E) Total cell number of spleen, liver, lung, MLN, and thymus of naïve mice. (F) Frequency of CD3+, (G) CD3+ CD8+, and (H) CD3+ CD4+ T cells from organs of mice as in (E). (I) Frequency of CD19+ B cells in spleen, lung, and MLN of naïve mice. (J) Frequency of double-positive and (K) double-negative T cells in the thymus of naïve mice. (L) Serum analysis of naïve mice. (M) Analysis of liver function in naïve mice. (N) Gating strategy for identifying cytokine-producing CD4+ T cells. (O) Frequency of IFN-γ, IL-4, IL-10, and IL-13-expressing CD4+ T cells. MLN cells from naïve mice were restimulated with PMA/Ionomycin in the presence of monensin, after which CD4+ T cells stained intracellularly for indicated cytokines. Results are representative of two independent experiments with 7–9 mice/group. Data are expressed as mean ± S.E.M. NS, P > 0.05; * P P P t test. Underlying data can be found in S1 Data. AST, aspartate aminotransferase; CD3, cluster of differentiation 3; CD4, cluster of differentiation 4; CD8, cluster of differentiation 8; CD19, cluster of differentiation 19; FMO, fluorescence minus one; Foxp3, forkhead box P3; FSC, forward scatter; IFN-γ, interferon gamma; IgE, immunoglobulin E; IL-4, interleukin-4; IL-10, interleukin-10; IL-13, interleukin-13; IL-4Rα, interleukin-4 receptor alpha; MLN, mesenteric lymph node; ND, not detectable; NS, not significant; PMA, phorbol myristate acetate; SSC-A, side scatter A; TNF-α, tumor necrosis factor alpha; Treg, regulatory T.