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

Fig. 3. HMGA2 is enriched in exosomes derived from EBV-positive NPC cells and increases vascular permeability and EndMT in endothelial cells in vitro.

Fig. 3

A HMGA2 protein levels in exosomes derived from EBV-negative NPC cell lines (HK1, HONE1, SUNE1, HNE1, CNE2, TW03 and 5-8F cells) and EBV-positive NPC cell lines (NPC43, C666-1 and C17 cells) were detected by Western blotting. Western blotting was performed at least three times. B Analysis of the HMGA2 content in corresponding cell-derived exosomes after overexpression and knockdown of HMGA2 in HK1 cells and NPC43 cells respectively. The change in HMGA2 expression in HUVECs treated as indicated for 12 h was analyzed by Western blotting (C) and immunofluorescence for HMGA2 (green), with DAPI (blue) staining of cell nuclei (D). Western blotting was conducted three times. E Representative images (high panel) and histogram of the quantification (low panel) of the endothelial cellular morphology alteration after the cells were treated as indicated for 48 h. ***p < 0.001. F HUVECs treated as indicated for 48 h were analyzed by Western blotting for the TJ proteins, CD31, α-SMA, Vimentin and S100A4. Western blotting was conducted three times. G The permeability of treated HUVEC monolayers grown on 0.4 μm filters was measured by the appearance of rhodamine-dextran. ***p < 0.001. H 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.