2.7. Flow cytometry
Cells from mouse midbrain or co culture system were stained by CD45, CD11c, CD11b, CD4, MHC Ⅱ, IFN- γ, IL-4 and IL-17A flow cytometry antibody (Biolegend). For IFN- γ, IL-4 and IL-17A staining, the cells were treated with 25 ng/ml PMA (Sigma Aldrich) and 1 mg/mL ionomycin (Sigma) for 5h before cell collection. After surface antibody staining, the cells were fixed and permeabilized with FIX &PERM MEDIUM A and FIX &PERM MEDIUM B (multi-Sciences), and then stained with intracellular factors antibody at 4 ℃. After the staining step, the cells were washed with PBS and analyzed by FCM.