Extended Data Figure 7 |. Rescued Treg cells in male Foxp3^LSL mice exert long-term control of adaptive and innate immune cells.

Mice were treated with a single dose of 4-OHT at 2 weeks of age and analyzed 4 months later. a, Body weight of rescued Foxp3^LSLCd4^creERT2 and control Foxp3^DTR-GFPCd4^creERT2 mice over a 4-month time course. Two-way ANOVA with Sidak’s multiple comparison test. b, Analysis of 4-OHT “functional” pharmacokinetics in 2-week-old male mice. Mice were injected with 4-OHT 3–72 hours before transfer of congenically marked recombination-proficient CD4 T cells from Cd4^creERT2R26^Tom mice. 4-OHT-induced recombination was assayed 7 days later by tdTomato expression among donor CD4 T cells as a readout for 4-OHT activity. Data are pooled from two independent experiments with 2 to 4 mice per time point each. Ctrl, no 4-OHT injection. c, Suppression of in vitro proliferation of conventional CD4 T cells induced by α-CD3 antibody and antigen-presenting cells by control Treg cells from Foxp3^DTR-GFP/y or rescued Treg cells from Foxp3^LSL/y mice. Two-way ANOVA with Tukey’s multiple comparison test. d-f, Numbers of activated, proliferating, and cytokine-producing conventional CD4 T cells (d), CD8 T cells (e) and myeloid populations (f) in indicated tissues. pLN, peripheral (brachial, axillary, and inguinal) lymph nodes; mLN, mesenteric lymph nodes; LP, lamina propria. Data are pooled from two independent experiments. Two-tailed t-tests with multiple hypothesis testing correction using the Holm-Sidak method. All error bars denote mean ± s.e.m. ns, non-significant; *, p<0.05; **, p<0.01; ***, p<0.001; ****, p<0.0001.