Fig. 1.

Analysis of the ApoE colocalization with αSN on vesicles from the human CSF. (A) αSN and identified apolipoproteins sequences’ coverages, and selected peptides spectra obtained after trypsin cleaved peptides’ fragmentation for each protein, for CSF samples after IP against αSN. Red, residues from a peptide domain observed at least once during the analysis; green, residues predicted to be difficult to observe by standard techniques; blue, residues found are chemically modified. (B) A codetection of ApoE and αSN using ELISA. Black bar, the detection of ApoE on anti-αSN antibody-bound complexes; gray bar, the detection of αSN on anti-ApoE antibody-bound complexes. (C) Levels of αSN bound to ApoE-positive vesicles in CSF from healthy controls compared with PD patients from the Stockholm cohort. Analysis was performed using the Mann–Whitney U test. P value < 0.05 was considered statistically significant. ****P < 0.0001. (D) The aggregation profile of the aggregation of ApoE alone (green), the aggregation of αSN alone (black), and the aggregation of αSN mixed with ApoE (blue). Each point represents the mean of 4 replicates, and errors bars represent SD of measurements. Comparison of the signal at plateau and the lag phase of aggregation between αSN with and without addition of the recombinant ApoE was performed using the Mann–Whitney U test. P value < 0.05 was considered statistically significant. (E) TEM images with the immunogold labeling representing αSN and ApoE colocalization on extracellular vesicles in the human CSF. (Scale bars, 50 nm.)