Figure 3

Dysregulated Treg percentage and function in Il2ra^−/−Tg mice. (a) Representative flow cytometry result of Treg cell percentage in CD4 single-positive T cells in thymus and pLN of Il2ra^−/−Tg and control mice. (b) Statistical analysis of Treg percentage and absolute number in thymus and pLN of Il2ra^−/−Tg (n=9), Il2ra^−/− (n=9), Il2ra^+/−Tg (n=9) and Il2ra^+/− (n=9) mice. (c) Expression of GITR, CTLA-4 and Helios of Treg cells in thymus and pLN from Il2ra^−/−Tg and littermate control mice. (d) Foxp3 expression as indicated by GFP MFI of Treg cells in thymus and pLN of Il2ra^−/−Tg (n=9), Il2ra^−/− (n=9), Il2ra^+/−Tg (n=9) and Il2ra^+/− (n=9) mice. (e) Percentage of CD69⁺ Treg cells in thymus and pLN of Il2ra^−/−Tg (n=5), Il2ra^−/− (n=5), Il2ra^+/−Tg (n=5) and Il2ra^+/− (n=5) mice. (f) CFSE-labeled CD45.1⁺CD4⁺Foxp3-GFP^-CD62L⁺ T cells were co-cultured with CD45.2⁺CD4⁺ Foxp3-GFP⁺ Treg cells from thymus and spleen/pLN of Il2ra^−/−Tg and Il2ra^+/− mice in the presence of soluble anti-CD3 (2 μg/ml) and anti-CD28 (1 μg/ml) and mitomycin C treated supporter cells for 72 h. Percentage of proliferated cells among live target cells were analyzed. (g) Ki-67 staining of Treg cells in thymus and pLN of Il2ra^−/−Tg (n=5), Il2ra^−/− (n=6), Il2ra^+/−Tg (n=5) and Il2ra^+/− (n=4) mice. (h) Annexin V staining of Treg cells in thymus and pLN of Il2ra^−/−Tg (n=7), Il2ra^−/− (n=7), Il2ra^+/−Tg (n=7) and Il2ra^+/− (n=7) mice. Data are shown in mean±S.E.M. *P<0.05, **P<0.01, ***P<0.001. (Student’s t test)