Fig. 1.

Normal development of non-pathogenic Th17 cells from Satb1-deficient T cells. a qPCR of CD4⁺ CD8⁺ (DP), CD4⁺ CD8⁻ (CD4SP), and CD4⁻CD8⁺ (CD8SP) thymocytes and CD8⁺, CD25⁻CD44^low CD4⁺, CD25^high CD4⁺, CD25⁻ CD44^high CD4⁺, and PPs eYFP⁺ CD4⁺ T cells for Satb1 mRNA expression. b Numbers of DP, CD4SP, and CD8SP cells in the thymus of 4-week-old Thpok^Cre Satb1^fl/fl and Thpok^Cre Satb1^wt/wt littermate controls. c Flow cytometry of CD4⁺ T cells from the LNs and PPs of Thpok^Cre Satb1^wt/wt and Thpok^Cre Satb1^fl/fl mice for intracellular IL-17 expression. The frequencies of IL-17⁺ in CD4⁺ T cells are shown. d Flow cytometry of splenic CD4⁺ T cells for intracellular IL-17 and IFN-γ expression. The frequencies of IL-17⁺ and IFN-γ⁺ in CD4⁺ T cells are shown. CD25⁻CD44^low CD4⁺ T cells (1 × 10⁶) from Thpok^Cre Satb1^wt/wt and Thpok^Cre Satb1^fl/fl mice were transferred to Rag2^−/− mice, and splenic CD4⁺ TCRβ⁺ T cells were analyzed on day 7 after transfer. e The frequencies of IFN-γ⁺ Th1, IL-4⁺ Th2, IL-17⁺ Th17 and Foxp3⁺ iTreg cells. CD25⁻CD44^low CD4⁺ T cells from Thpok^CreSatb1^wt/wt and Thpok^CreSatb1^fl/fl mice were cultured under Th1, Th2, Th17, and iTreg conditions for 3 days. Each symbol represents an individual mouse, and the horizontal lines indicate the mean values. f Flow cytometry of CD4⁺ T cells from the draining LNs of Thpok^Cre Satb1^wt/wt and Thpok^Cre Satb1^fl/fl mice immunized with MOG-CFA, staining for CD4 and intracellular IL-17 on day 7 after immunization. The frequencies of IL-17⁺ in CD4⁺ T cells are shown. The bar graphs (a–d, f) show the mean ± s.d. (n = 3). The results are representative of three independent experiments (a–f). *P < 0.05, ***P < 0.0001 (two-tailed unpaired Student’s t-test)