Fig. 6

STIM1 and STIM2 control the differentiation and trafficking of tissue-resident Treg cells. a Analysis of intravascular and tissue-resident Foxp3⁺ Treg cells in the parenchyma of BM, liver, and lung of male WT and Stim1/2^Foxp3 mice by flow cytometry; means of 10–13 mice. For the gating strategy of tissue-resident Treg cells see Supplementary Figure 7i. b Analysis of WT and Stim1/2-deficient Treg cells in LNs and the parenchyma of BM, liver, and lung of female heterozygous WT and Stim1/2^Foxp3 mice by flow cytometry. Data is representative of three mice; for the gating strategy of tissue-resident Treg cells see Supplementary Figure 7i. c Generation of mixed BM chimeric mice using T cell-depleted BM from male CD45.1⁺ WT mice and BM from male CD45.2⁺ Stim1/2^Foxp3 mice at a 1:1 ratio. d Analysis of tissue-resident Treg cells in mixed BM chimeras. Distribution of CD45.1⁺ WT and CD45.2⁺ Stim1/2-deficient Treg cells in the thymus, spleen, and LNs, as well as the parenchyma of BM, liver, and lung analyzed by flow cytometry; means ± SEM of 4–6 mice. e–g Expression of tissue-homing receptors on Treg cells depends on SOCE. e Canonical pathway analysis (KEGG) based on differentially expressed genes (DEG) in anti-CD3/CD28 stimulated WT versus Stim1/2-deficient Treg cells. f Selected gene set enrichment analyses (GSEA) of DEG in WT versus Stim1/2-deficient Treg cells. g Heatmap analysis of DEG (>1.5 fold) encoding chemokine receptors (upper panel) and integrins (lower panel) in WT and Stim1/2-deficient Treg cells. Statistical analysis in a and d by unpaired Student’s t-test. *p < 0.05; **p < 0.01; ***p < 0.001