Extended Data Figure 4. Features of T-bet⁺ Treg cells.

a, T cell activation, CXCR3 expression, and cytokine production in 12 wk old control Foxp3^YFP-CreTbx21^WT/WT and Foxp3^YFP-CreTbx21^FL/WT (white circles) and experimental Foxp3^YFP-CreTbx21^FL/FL (black circles) mice. Bars, mean±s.e.m. Two-tailed t test (* denotes p value < 0.05; NS – not significant). Data is representative of 3 experiments, n ≥ 7 mice per group. b, Cumulative distribution function plot of the 561 genes up in Thy1.1⁺ CD44^hiRFP^hiCXCR3⁺ vs. CD44^hiRFP⁻ cells in Tbx21^RFP-CreERT2 mice compared to all genes differentially expressed in CD4⁺CD25⁺ Treg vs. CD4⁺CD25^lo exTreg cells. P=0.2xE⁻¹⁵, two-sample Kolmogorov-Smirnov test. c, Expression of CCR5 (above) and CD29 (below) in CD44^loCD62L^hi naïve (blue histogram), CD44^hiCXCR3⁻ (black histogram) and CD44^hiCXCR3⁺ (red histogram) Treg (left) and CD4⁺Foxp3⁻ (right) T cells from spleens of Foxp3^YFP-CreTbx21^WT/WT mice. d, Expression of CXCR3 (left), CCR5 (middle), and CD29 (right), gated on CD4 T cells in spleens of Foxp3^YFP-CreTbx21^WT/WT and Foxp3^YFP-CreTbx21^FL/FL mice. e, Dendrogram represents cluster analysis of TCR sequences in CD44^hiCXCR3⁺ (red symbols) and CD44^hiCXCR3⁻ (black symbols) Treg (right) and effector CD4 T (left) cells in spleens (white symbols) and lymph nodes (gray symbols) of DO11.10 TCRβ⁺ Tcra^+/− Foxp3 reporter mice. Sample preparation and statistical analyses are described in the Methods section. Pearson correlation of clonotype frequencies for the shared TCR clones was used for the generation of the dendrogram.