FIGURE 6:

K5 displaces p120 from VE-cadherin. K5-FLAG was expressed in primary dermal microvascular endothelial cells by adenoviral transduction. Cells were treated with vehicle (bottom) or 100 μM chloroquine (top). (A) After 24 h, cells were fixed and immunostained for VE-cadherin, p120, and FLAG as indicated. (B) The density of VE-cadherin–containing vesicles in individual cells was quantified. Thick line, median; boxes, interquartile range; whiskers, 90% range (20–22 cells/group); p < 0.001. (C) Individual vesicles were selected from K5-expressing chloroquine-treated cells, and the amount of VE-cadherin and p120 fluorescence signals was quantified. Most vesicles contain high levels of VE-cadherin fluorescence or high levels of p120 fluorescence but not both. Bars, 20 μm (main image), 80 μm (inset).