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. 2016 Jun 16;1(9):e86462. doi: 10.1172/jci.insight.86462

Figure 4. FTY720 treatment in vivo downregulated Th17 expression in the spleen of S1PR1(S5A) EAE mice.

Figure 4

MOG35-55-immunized C57BL/6J (WT) and S1PR1(S5A) EAE mice were treated with vehicle (1% cyclodextrin in PBS) or FTY720 (0.5 mg/kg) by daily i.p. injections up to day 8 after immunization. Splenocytes were harvested, and the total splenocyte counts were quantified by hemocytometer (A). Percentages of CD3+ (B), CD4+ (C), CD8+ (D), and CD11b+ (E) cells were quantified by flow cytometry. Splenocytes were also stimulated with PMA (50 ng/ml) and ionomycin (500 ng/ml) for 4 hours, and intracellular staining was performed to measure the expression of IL-17A, IFN-γ, and FOXP3 among CD4+ cells (F–I). Data represent 2 independent experiments (n = 4, mean ± SEM, *P < 0.05; **P < 0.01, ANOVA with Tukey’s multiple comparison test).