Skip to main content
. 2022 Apr 6;29(10):1439–1451. doi: 10.1038/s41417-022-00453-6

Fig. 1. Exosomes derived from EBV-positive NPC cells reduce endothelial cell TJ protein expression and promote EndMT.

Fig. 1

A Confocal microscopy analysis of HUVECs treated with PKH-67 (green)-labeled purified NPC cell-derived exosomes or PKH-67 wash buffer (scale bar = 20 μm). B The permeability of treated HUVEC monolayers grown on 0.4 μm filters was measured by the appearance of rhodamine-dextran, which was added to the top well at the beginning of the experiment, in the bottom well during a 2 h time course. The absorbance at 590 nm at each time point is indicated. ***p < 0.001. C HUVEC monolayers grown on 3 μm filters were treated as indicated before GFP-labeled 5-8F cells were seeded in the Transwell inserts. After 10 h, the GFP+ cells on the bottom side of the filters were quantified under a fluorescence microscope. ***p < 0.001. D Representative images (left panel) and histogram of the quantification (right panel) of endothelial cellular morphology alterations after the cells were treated with PBS, HK1-Exo, CNE2-Exo, NPC43-Exo, C666-1-Exo and C17-Exo. ***p < 0.001. E HUVECs treated with PBS, HK1-Exo, CNE2-Exo, NPC43-Exo, C666-1-Exo and C17-Exo were analyzed by Western blotting for TJ proteins (ZO-1, VE-cadherin, occludin and claudin-5), CD31, α-SMA, Vimentin and S100A4. F HUVEC monolayers were treated as indicated for 48 h and analyzed by immunofluorescence for ZO-1 (green), VE-cadherin (green), occludin (green) and claudin-5 (green), CD31 (red), α-SMA (green), Vimentin (red) and S100A4 (green). DAPI (blue): cell nuclei.