Figure 2. Integrin β3 Expression on Th17 Cells in EAE.

CD4⁺ T cells from dLNs and CNS were analyzed by flow cytometry during active EAE (day 12–16) induced in IL-17Cre-YFP fate-tracking mice.

(A) Representative flow cytometry plots of integrin β3 and intracellular IL-17 staining following PMA/ionomycin stimulation in live CD4⁺ cells from indicated sites.

(B) IL-17 and integrin β3 co-expression in live CD4⁺ T cells from naive and EAE LNs, stimulated as indicated (all EAE plots are from same donor for comparison).

(C) EAE LN cells were stimulated with PMA/ionomycin, and live CD4⁺ cells were then gated according to high or low integrin β3 expression to determine expression of intracellular IL-17 protein and IL17Cre-mediated YFP.

(D) Histogram comparing RORγt expression in integrin β3^hi and integrin β3^lo CD4⁺ EAE LN cells.

(E) EAE LN cells stimulated with PMA/ionomycin and analyzed for expression of IL-17 and RORγt in integrin β3^hi and integrin β3^lo CD4⁺ cells.

(F) Representative plots of RORγt⁺integrin β3^hi population within live CD4⁺ cells from EAE LNs and naive LNs.

(G) Percentage of integrin β3^hi cells gated from live CD4⁺ cells, with data pooled from three separate experiments.

Data are representative of at least three separate experiments, with two to four mice per group. Statistical significance was analyzed by one-way ANOVA. Error bars indicate mean ± SD.