Fig. 4

SENP3 regulates Treg cell effector programs and stability. a–e Splenic Treg cells (CD4⁺CD25⁺YFP⁺) obtained from 6-week-old female Senp3^+/+Foxp3-Cre (WT) mice and Senp3^fl/flFoxp3-Cre (KO) were stimulated with anti-CD3 and anti-CD28 antibodies for 24 h and subjected to RNA sequencing. A heat map of highly variable genes (top 1000) (a) and a heat map of genes with a fold change >2 and adjusted P value < 0.01 (b) were shown. Scatter plot showing gene expression in the KO versus WT Treg cells (c). Teff cell-specific genes are highlighted in blue, and the Treg cell-specific genes are highlighted in red. Clustering of upregulated Teff cell-specific genes identified in SENP3-deficient Treg cells relative to that in Senp3-WT Treg cells (d). Treg cell-specific gene expression in SENP3-deficient Treg cells and Senp3-WT Treg cells (e). f qRT-PCR analysis of the indicated genes in anti-CD3 and anti-CD28 antibody-stimulated Treg cells from Senp3^+/+Foxp3-Cre and Senp3^fl/flFoxp3-Cre mice. g, h Flow cytometric analysis of Foxp3 expression (g, the numbers above the graphs indicate MFI of Foxp3) or IFN-γ and IL-17 expression (h) in Senp3^+/+Foxp3-Cre and Senp3^fl/flFoxp3-Cre Treg cells (CD4⁺CD25⁺YFP⁺) stimulated with anti-CD3 and anti-CD28 antibodies for 24 h. Data are presented as representative plots (left) and summary graphs of percentage and MFI of Foxp3 (right). Data in f, g, h are representative of three independent experiments and are presented as the means ± SEM. n = 5. Two-tailed unpaired Student’s t tests were performed. *P < 0.05; **P < 0.01