Figure 6. Scinderin accounts for defective function of CYLD^−/− Treg cells.

Analysis of Cyld^fl/fl Scin^fl/fl Foxp3^cre mice. A and B, The expression of CD62L and CD44 of splenic CD4⁺ T cells was assessed by flow cytometry. C-E, The expression of IL-4 was determined qPCR (C) and flow cytometry (D and E). Splenic YFP⁺ Treg cells were sorted for qPCR. For flow cytometry, lung CD4⁺ T cells were activated with PMA and ionomycin for 3 hrs prior to intracellular staining of IL-4 and Foxp3 (D) and the absolute number of IL-4⁺ Foxp3⁺ and Foxp3⁻ T cells were calculated (E). F and G, histological analysis of lungs from Cyld^fl/fl Scin^fl/fl Foxp3^cre double KO mice. The lung sections of 2 month-old mice were stained with H&E (F) and the clinical scores were assessed (G). H and I, N. Brasiliensis infection in Cyld^fl/fl Scin^fl/fl Foxp3^cre double KO mouse. H, The number of worms in the intestines was examined at day 7 after infection. I, The recruitment of eosinophils to the peritoneum was assessed. Representative of at least two independent experiments are shown. Combined plots for B (n=6), C (n=4~6), E (n=3~5), G (n=3~6), H (n=9~11) and I (n=9~11) are shown. Error bars indicate SEM. ∗ P<0.05, ∗∗ P<0.01, ∗∗∗∗ P<0.0001 (Mann-Whitney test)