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. 2021 Jun 10;9:689947. doi: 10.3389/fcell.2021.689947

FIGURE 2.

FIGURE 2

Exosomes derived from Clear cell renal cell carcinoma cells increase the permeability of the endothelial cells. (A) Western blot analysis of Vimentin, β-catenin, E-cadherin, N-cadherin, Zo-1, Claudin expression in HUVECs incubated with CM of 786-O and 786-O-SR. (B) Transmission electron microscopy of exosomes derived from 786-O and 786-O-SR. Scale bar, 100 nm. (C) Western blotting analysis of CD81 and TSG101 in 786-O, 786-O-SR and their exosomes. (D) Nanoparticle tracking analysis of the size distribution and median diameter of particles per μg exosomes from 786-O and 786-O-SR. (E) The presence of BODIPY TR ceramide fluorescence in HUVECs after adding dye-labeled exosomes derived from 786-O and 786-O-SR cells for 48 h. HUVECs incubated with PBS were used as a negative control. Red: BODIPY TR ceramide; Green: Calcein AM; Blue: DAPI. (F) Western blot analysis of β-catenin, Vimentin, E-cadherin, N-cadherin, Zo-1, Claudin expression in HUVECs incubated with exosomes of 786-O and 786-O-SR. (G) Transendothelial invasion assay analysis of the number of GFP-expressing 786-O cells that invaded through HUVECs monolayers cultured with exosome derived from 786-O or 786-O-SR. Mean ± SEM are provided (n = 3). **P < 0.01, ***P < 0.001, according to two-tailed Student’s t-test. exo, exosomes; NC, negative control.