FIGURE 1.

The number of Tregs is increased in the spleen of CYLD-deficient mice. A, wild type and CYLD-deficient CD4⁺ cells from splenocytes were analyzed for Foxp3 expression by flow cytometry. APC, allophycocyanin; PE, phycoerythrin. B, graph showing the mean and distribution of CD4⁺ Foxp3⁺ splenic T cells from wild type and CYLD-deficient mice. Twelve mice per group were analyzed. **, p < 0.01. C and D, FACS staining of mesenteric lymph nodes (MLN) performed as in A and B. E and F, FACS staining of thymocytes performed as in A and B. *, p < 0.05. Error bars represent the mean ± S.D. p values were calculated by using Student's t test. G and H, proliferative and suppressive function of CYLD-deficient CD4⁺ T cells. Wild type or CYLD-deficient CD4⁺CD25⁻ cells and CD4⁺CD25⁺ cells, or CD4⁺CD25⁻ cells co-cultured with CD4⁺CD25⁺ cells, were stimulated with antigen-presenting cells and anti-CD3. Cultures were incubated for 72 h and pulsed with [³H]TdR for the last 6 h of culture. One representative experiment of two separate experiments is shown.