Fig. 4

Analysis of α-syn co-occurrence with trafficking markers. Representative confocal micrographs of α-syn monomer treated pBECs on filters with α-syn added for 10 min to the luminal side (a). Green show α-syn monomer stain, blue is Hoechst stain and red is the EEA1 stain. Lower right micrograph shows the outcome of IMARIS spot segmentation and colocalization analysis between α-syn and EEA1 channels. Scale bars show 15 µm. b Semi-quantification of α-syn co-occurrence with different compartments in pBEC: EEA1 (early endosome), RAB7 (endosome-lysosome), VPS35 (endosome-golgi, retromer), Rab8a (Golgi-plasma membrane), Caveolin1 (Caveolae/transendothelial channels) and Clathrin (endo-lysosomal vesicles). Statistical difference was tested using an ordinary two-way ANOVA followed by Tukey's multiple comparisons test