Supplementary Figure 3: The EAE phenotype of adoptive transfer of sorted CD4+ T cells from Ascf/+Lck-Cre and Ascf/–Lck-Cre mice.
From: T cell–intrinsic ASC critically promotes TH17-mediated experimental autoimmune encephalomyelitis
(a) Absolute numbers of CD4+ IL-17A+ cells (left panel) and concentrations of IL-17A in the supernatant of cells (right panel) from the draining lymph nodes and spleens of Ascf/+Lck-Cre and Ascf/-Lck-Cre mice immunized with MOG35-55 and harvested 10 days after immunization, followed by ex vivo re-stimulation in the presence of MOG35-55 and IL-23. (b-d) Cells from lymph nodes of Ascf/+Lck-Cre and Ascf/-Lck-Cre mice 10 days after immunization with MOG35-55 were re-stimulated with MOG 35-55 in vitro in the presence of recombinant IL-23 for 5 days, sorted for CD4+ T cells by flow cytometry, and then transferred into CD45.1 congenic mice. (b) Graph represents the average clinical score after T cell transfer. (c) Absolute numbers of CNS-infiltrating CD45.2+ CD4+ cells were determined at the peak of disease. Brains and spinal cords were harvested together and CD45.2+CD4+ cells were stained with anti-CD45.2 and anti-CD4 antibodies, followed by flow cytometric analysis. (d) Absolute numbers of CNS-infiltrating cells were determined at the peak of disease. Brains and spinal cords were harvested together and mononuclear infiltrating cells were stained with anti-CD45, anti-CD4, anti-CD8, anti-F4/80, anti-Ly6C, anti-CD19 and anti-Ly6G antibodies, followed by flow cytometric analysis. Two-way ANOVA (b) and Unpaired t test (a, c and d) were used to analyzed the data. Data are representative of three independent experiments (a-d). n=5 mice per group in each experiment. Error bars represent s.e.m. *P < 0.05 (a, c and d). Error bars represent s.d. *P < 0.05 (b).
