Figure 3. miR-155 is required for inflammatory T cell development during the induction phase of EAE.

Mir155^+/+ and Mir155^−/− mice were harvested 13 days following immunization with MOG_35–55. A. Mononuclear cells were purified from Mir155^+/+ and Mir155^−/− brains and intracellular staining was conducted to identify CD4⁺ lymphocytes producing IL-17A and/or IFN-γ (n=5). B. Total numbers of Th17 and Th1 cells in the brain, in addition to the percentage of Th17 and Th1 cells among total CD4⁺ T cells is shown on the right (n=5). C. WT and Mir155^−/− splenocytes were analyzed for expression of BIC, IL-17A and IL-23 p19 mRNA by qPCR (n=5) and D. intracellular staining was used to determine the number of Th17 and Th1 cells (n=5). E. Splenocytes were restimulated with MOG_35–55 and E. proliferation was assayed by ³[H] thymidine incorporation (n=5) and F. the production of IL-17A, IFN-γ, IL-6 and GM-CSF measured by ELISA (n=5). G. Expression of BIC and IL-17A mRNA in the LNs was assayed by qPCR (n=5) and H. the number of Th17 and Th1 cells was also quantified by flow cytometry (n=5). Error bars represent +/−SEM and * denotes statistical significance with a p value of <0.05 according to a student’s two-tailed t-test. +/+ = Mir155^+/+; −/− = Mir155^−/−. See also Figure S3.