Fig. 11.

sEV-dependent α-Syn transmission in primary microglia. a. α-Syn pre-formed fibrils (PFF) were pre-incubated with sEVs derived from SNCA KO mouse brains (PFF + sEVs), for 20 h at 37 ^oC. Microglia cells were treated with PFFs or sEVs alone or PFF + sEVs for 2 h. Uptake of α-Syn and sEVs were monitored 2 h and 6 h post-addition. Cells were fixed and immunostained against α-Syn (D10, light gray) and Iba1 (green). Cell nuclei were stained with DAPI (blue). Representative Imaris images show the uptake of α-Syn and sEVs in all three conditions (PFFs, sEVs, and PFF + sEVs) 2 h and 6 h post-treatment. Arrowheads depict PFF colocalization with sEVs. Colocalization of α-Syn with sEVs is depicted in magenta. Scale bar 10 μm. Graphs present the total volume of α-Syn per cell (b), the mean volume of α-Syn individual puncta (c), the total volume of sEVs per cell (d), and the mean volume of sEV puncta (e) under the different conditions. Data are presented as the mean ± SEM of minimum three independent cell preparations, with at least two replicates per assay; one-way ANOVA with Tukey’s correction was used for (c) and (e), two-way ANOVA with Tukey’s correction for (d), and multiple t test for (b). Statistical significance was set as *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001