Fig. 3

α-Synuclein in CNS was drained to the macrophage and activated it in the dCLN through meningeal lymphatics. A Representative fluorescence images of dCLNs of C57 mice injected (i.c.m) with vehicle or AF647-α-synuclein (red). Scale bar, 200 μm. B Number of macrophages containing AF647-α-synuclein in dCLNs of C57 mice injected (i.c.m) with vehicle or AF647-α-synuclein was evaluated by flow cytometry. The macrophages were labelled by CD11b-FITC and CD45-PE-Cy7. C Quantitative analysis of the number of macrophages containing AF647-α-synuclein in dCLNs of C57 mice. D Representative fluorescence images of dCLNs in WT and A53T mice. α-Synuclein in dCLN was immunolabelled by MJFR-1 (red). Scale bar, 200 μm. E Macrophages in dCLN were sorted by FACS. The macrophages were labelled by CD11b-FITC and CD45-PE-Cy7. F Quantitative analysis of α-synuclein oligomer in macrophages of dCLNs and total dCLNs using MSD. N = 3 independent experiments in each group. G Quantitative analysis mRNA levels of Il1b, Il6 and Tnf in BMDMs treated with vehicle control, 100 nM α-syn monomer, or 100 nM α-syn oligomer using qPCR. N = 3 independent experiments in each group. H Quantitative analysis of the levels of IL-1β, IL6 and TNF-α using ELISA, released by BMDMs treated with vehicle control, α-syn monomer, or α-syn oligomer. N = 3 independent sample pools in each group. Values are means ± S.E.M, t test (C, F), one-way ANOVA test (G, H). *, P < 0.05; **, P < 0.01; ****, P < 0.0001