Figure 9.

Cdc42-deficient nTreg cells from Cdc42^fl/flFoxp3^YFP-Cre/+ mice and from Cdc42^fl/flFoxp3^YFP-Cre bone marrow chimeric mice are impaired in competitiveness and stability. (A) The ratio of Foxp3⁺YFP⁺ vs Foxp3⁺YFP⁻ cells from the spleen of Cdc42^+/+Foxp3^YFP-Cre/+ and Cdc42^fl/fl Foxp3^YFP-Cre/+ mice. (B) Percentages of BrdU-incorporated cells among Foxp3⁺YFP⁺ cells from the spleen of Cdc42^+/+Foxp3^YFP-Cre/+ and Cdc42^fl/flFoxp3^YFP-Cre/+ mice. (C) Mean fluorescence intensity (MFI) of active caspase 3 among Foxp3⁺YFP⁺ cells from the spleen of Cdc42^+/+Foxp3^YFP-Cre/+ and Cdc42^fl/flFoxp3^YFP-Cre/+ mice. (D) Representative flow cytogram of staining of IL-17, IFN-γ and IL-4 in CD4⁺Foxp3⁺YFP⁻ and CD4⁺Foxp3⁺YFP⁺ cells from the spleen of Cdc42^fl/flFoxp3^YFP-Cre/+ mice. (E) Expression of Foxp3 (MFI) in Foxp3⁺YFP⁻ and Foxp3⁺YFP⁺ cells from the spleen of Cdc42^fl/flFoxp3^YFP-Cre/+ mice. (F) Expression of the indicated Treg functional markers on Foxp3⁺YFP⁻ and Foxp3⁺YFP⁺ cells from the spleen of Cdc42^fl/flFoxp3^YFP-Cre/+ mice. Numbers indicate MFI. (G) Bone marrow cells from Cdc42^+/+Foxp3^YFP-Cre or Cdc42^fl/fl Foxp3^YFP-Cre mice were mixed with bone marrow cells from BoyJ mice at 1:1 ratio and transplanted into lethally irradiated BoyJ mice. Eight weeks later, donor-derived (CD45.2⁺) CD4⁺Foxp3⁺ cells in the spleen of recipient mice were analyzed for percentages of total CD4⁺Foxp3⁺ cells and CD4⁺Foxp3⁺ cells that express IFN-γ or IL-4. n = 4 or 5. Error bars indicate SD. *p < 0.05. **p < 0.01.