Fig. 5
Proteome analysis of primary choroid plexus epithelial (CPE)-derived extracellular vesicles (EVs) and complement activation in the CP after stimulation with Aβ oligomers (AβO). (a, b) Venn diagrams showing overlap of the total number of proteins identified in EVs separated from the apical medium of primary CPE cells after 2 h of stimulation with (a) scrambled peptide (green) or (b) AβO (pink) (n = 3). (c) Venn diagram showing overlap of proteins identified in at least two out of three replicates of EVs separated from the apical medium of primary CPE cells after 2 h of stimulation with scrambled peptide (green) or AβO (pink) and 24 h of incubation (n = 3). (d) Ingenuity pathway analysis (IPA) of differentially regulated proteins in EVs separated from the apical medium of primary CPE cells after 2 h of stimulation with AβO or scrambled peptide and 24 h of incubation (n = 3). (e) Heat map of z-scores (calculated from log2-transformed label-free quantification (LFQ) protein intensities) for proteins differentially expressed (P < 0.01) in EVs separated from the apical medium of primary CPE cells after 2 h of stimulation with AβO or scrambled peptide and 24 h of incubation (n = 3). Proteins are ranked in ascending order according to their fold change value for the AβO versus the scrambled group. (f) qRT-PCR analysis of the complement component 3 (C3) in the CP 6 h after the icv injection of scrambled peptide (black) or AβO (grey) in C57BL/6J mice (n = 5), analyzed using an unpaired t-test. (g) 3D reconstructions of representative confocal images of C3 (red) in the CP 6 h after the icv injection of AβO or scrambled peptide in C57BL/6J mice (n = 3). Cell nuclei are counterstained with Hoechst (blue). Scale bar represents 100 µm