Figure 5.

VE-cadherin required for Ang-1-induced cell adhesion to extracellular matrix. (A) Quantification of relative cell adhesion to gelatin-coated substrates of BAECs transfected with siRNA against VE-cadherin (siVE-cadherin), Rap1 (siRap1), or control siRNA (siCT), as indicated. Adhesion assays were performed in absence or presence of Ang-1 (100 ng/mL). Results shown as relative ratio to untreated and siCT-transfected cells. (B) Representative confocal micrographs of BAECs stained for total phosphotyrosine (4G10; green) and transfected with siCT, siVE-cadherin, or siRap1. Where indicated, cells were stimulated with Ang-1 (100 ng/mL) for 30 min prior to fixation. Arrows point towards cell membrane where phosphotyrosine staining was strongest. Scale bar: 20 μm. (C) Representative confocal micrographs images of BAECs transfected with GFP-FAK plasmids and transfected with siCT, siVE-cadherin, or siRap1. Where indicated, cells were stimulated with Ang-1 (100 ng/mL) for 30 min prior to fixation. (D,E) Quantification of number of focal adhesions (FAs) per cell and FA size in cells shown in C. Total of five cells per condition were counted. Graphs are representative of three independent experiments each yielding similar results; * p < 0.05.