Figure 2.

Homozygous RhoA deletion in Treg cells dampens Treg cell homeostasis and induces Treg cell plasticity. (A) Left, representative flow cytogram of Foxp3 staining in CD4⁺ cells from the spleen of RhoA^+/+Foxp3^YFP-Cre and RhoA^Flox/FloxFoxp3^YFP-Cre mice. The numbers indicate percentages of CD4⁺Foxp3⁺ Treg cells. Middle, average percentages of CD4⁺Foxp3⁺ Treg cells. Right, average numbers of CD4⁺Foxp3⁺ Treg cells. (B) Treg cell proliferation. Percentages of CD4⁺Foxp3⁺ Treg cells incorporated with BrdU are shown. (C) Treg cell apoptosis. The expression levels (MFI: Mean fluorescence intensity) of active caspase 3 in CD4⁺Foxp3⁺ Treg cells are shown. (D) The expression levels of Foxp3 in Treg cells. (E) Left, representative flow cytogram of IL-17, IFN-γ, and IL-4 staining in CD4⁺Foxp3⁺ Treg cells. The numbers indicate percentages of IL-17⁺, IFN-γ⁺, and IL-4⁺ Treg cells. Right, average percentages of IL-17⁺, IFN-γ⁺, and IL-4⁺ Treg cells. (F) Left, representative flow cytogram of RORγT, T-bet and GATA3 staining in CD4⁺Foxp3⁺ Treg cells. The numbers indicate percentages of RORγT ⁺, T-bet⁺, and GATA3⁺ Treg cells. Right, average percentages of RORγT ⁺, T-bet⁺, and GATA3⁺ Treg cells. (G) Left, representative histogram of the expression levels of CTLA-4, GITR and PD-1 in CD4⁺Foxp3⁺ Treg cells. The numbers above the graphs indicate MFI. Right, average MFI of CTLA-4, GITR and PD-1 in CD4⁺Foxp3⁺ Treg cells. n = 3 mice. Data are representative of two independent experiments. Error bars indicate SD. **p < 0.01.