FIGURE 6.

An intact ultrastructure is necessary for EV‐induced modification to αsyn fibrillization. The effect of treating sEVs with methanol alone (MeOH), sarkosyl in addition to methanol (MeOH/Sark), or left untreated (Untreat) to their ability to alter αsyn fibrillization was assessed by: (a) TEM, where red arrors indicate the detection of intact sEVs following PMCA in the untreated group, (b) ThT on protein exposed to PMCA for 12 h, where values obtained for each experimental replicate represented the average fluorescence of triplicate wells after subtraction of a blank well to account for background fluorescence (R.F.U = relative fluorescent units). Data presented as mean±SEM (n = 3), (c) western immunoblot in the presence of PK, (d) western immunoblot on samples separated to soluble and insoluble fractions and (e) analytical centrifugation. Shown are enhanced van Holde‐Weischet integral distributions from extrapolation of sedimentation velocity data displayed in Fig. S1D–F. Apparent sedimentation coefficient distributions are plotted as a function of boundary fraction for αsyn in the presence of untreated EVs (upright black triangles, red fill), MeOH‐treated EVs (upright coral red triangles, white fill), or MeOH/Sark‐treated EVs (inverted light‐coral red triangles, white fill). (f) Coomassie of untreated sEVs and those chemically modified with methanol alone or methanol or sarkosyl