Fig. 1

Schematic diagram of the experimental design. Foxp3-RFP mice were immunized with 15 or 100 μg of recombinant human α-syn in complete Freund’s adjuvant and re-immunized after 4 weeks with the same amount of protein in incomplete Freund’s adjuvant. Five days later, the animals were killed and the lymph nodes and brain dissected for analysis. Blood was obtained to determine antibody load in serum. A single lymph node cell suspension was made and either analyzed by flow cytometry or incubated for 6 h, after which the cells were lysed for analysis by Western blot. Microglia were isolated from brains and analyzed by flow cytometry. Alternatively, the animals were perfused at the time of death and brains processed for immunohistochemistry