Fig. 3

STIM1 and STIM2 control a complex transcriptional network in Treg cells. a–f Gene expression analysis of WT and Stim1/2-deficient Treg cells isolated from female heterozygous Stim1/2^Foxp3 mice by RNA-sequencing. The isolation strategy of Treg cells is shown in Supplementary Figure 2a. a MA plots of differentially (>1.5-fold) expressed genes (DEG) in unstimulated WT and Stim1/2-deficient Treg cells and Treg cells stimulated with anti-CD3/CD28 for 16 h. Genes significantly (p < 0.05) upregulated and downregulated are depicted in red and blue, respectively. b Pathway and network analysis based on DEG in unstimulated WT and Stim1/2-deficient Treg cells. Downregulated and upregulated pathways are shown in blue and red, respectively. c Volcano plot of DEG (gray) between unstimulated WT and Stim1/2-deficient Treg cells overlaid with upregulated (red) or downregulated (blue) genes of the ‘IL-2/STAT5 signaling signature’³⁵. d Volcano plot of DEG (gray) between unstimulated WT and Stim1/2-deficient Treg cells overlaid with upregulated (red) or downregulated (blue) genes of the ‘common Treg signature’⁴. e Heatmap analysis of selected DEG (>1.5-fold) encoding regulatory molecules (upper panel) and cell surface receptors (lower panel) in WT and Stim1/2-deficient Treg cells. f Heatmap of selected differentially expressed (>1.5 fold) transcription factors in WT and Stim1/2-deficient Treg cells. g Analysis of protein expression of DEG on conventional CD4⁺ T cells (Tcon), WT and Stim1/2-deficient Treg cells by flow cytometry. Bar graphs represent the means ± SEM of four mice. Statistical analysis in (g) by unpaired Student’s t-test. *p < 0.05; **p < 0.01; ***p < 0.001