Fig. 2

DU145 and DU145R80-derived LO protein cargo. a-b Western blot analysis for the indicated proteins on iodixanol purified fractions of large EVs from both cell lines. Fractions enriched in LO are indicated in red. Whole cell lysates from DU145R80 used as positive control. c αV-integrin expression on LO145 (left panels) and LOR80 (right panels), purified by iodixanol gradient (1.15 g/mL fractions), measured by Amnis® Imaging Flow Cytometer upon labeling with a specific PE-conjugated antibody. Representative images were acquired at 40× magnification (bar size 10 μm). d Flow cytometric analysis of both LO145 and LOR80 purified by iodixanol gradient (1.15 g/mL fractions), identified as intact vesicles in a range size between 1 and 10 μm and positive for αV-integrin. e Flow cytometric analysis of LO145 and LOR80 purified by iodixanol gradient (1.15 g/mL fractions) and positive (red) for CK18 (upper panel) and αV-integrin (lower panel) as compared to negative controls (black)