Figure 6. TSC1-deficient Treg cells acquire a Th17-like phenotype in female Foxp3^YFPCre/+Tsc1^f/f mice.

(A) Flow cytometric analysis of YFP, CD25, and Foxp3 expression in CD4⁺ T cells from spleen of female Foxp3^YFPCre/+Tsc1^f/f mice. (B) Sorted CD4⁺CD25⁺NRP1⁺YFP^– or CD4⁺CD25⁺NRP1⁺YFP⁺ Treg cells from spleen of female Foxp3^YFPCre/+Tsc1^f/f mice were stimulated with anti-CD3/CD28 for 36 hours. Cytokine production was measured by Bio-Plex multicytokine assay. (C) Flow cytometric analysis of IL-17 production by CD4⁺Foxp3⁺YFP^– or CD4⁺Foxp3⁺YFP⁺ Treg cells from MLNs, Peyer’s patch (PP), and cLP of female Foxp3^YFPCre/+Tsc1^f/f mice. Cells were restimulated with PMA/ionomycin for 6 hours. Data are representative of (A and C) or compiled from (B) three to four independent experiments. Error bars indicate the mean ± SD. *P < 0.05 and ***P < 0.001 by two-tailed, unpaired Student’s t test.