FIGURE 5. RhoA specific inhibitor Y16 blocks Th17 differentiation.

(A) Purified naïve CD4⁺ T cells (1 × 10⁶/mL) pooled from 8 WT mice were stimulated with plate-bound anti-CD3 plus free anti-CD28 for 2 days without or with Y16 (0~50 μM). IL-17 and IL-21 in the culture supernatants were determined by ELISA. (B-D) Naïve CD4⁺ T cells were differentiated under Th17 conditions for 4 days and restimulated with PMA plus ionomycin for 5 h, in the presence of vehicle (Veh) or Y16 (30 μM) throughout the culture. Cells were collected for IL-17/IFN-γ intracellular staining. Percentages of IL-17⁺ or IFN-γ⁺ CD4⁺ T cells are shown in representative dot plots (B) and summarized in a bar graph (C). IL-17 and IL-21 in the culture supernatants were determined by ELISA (D). Data are representative of 2 independent experiments. **P < 0.01 compared to vehicle-treated groups