Fig 5.

Overexpression of integrin αv and β3 subunits increases adhesion and affects migration of endothelial cells. (A) Total cell extracts of HUVEC expressing either GFP (control) or integrin αv and β3 subunits were analyzed by Western blotting using specific antibodies. (B) HUVEC as described in panel A were plated on vitronectin, and adhesion was measured as described in Materials and Methods. Results are given as OD values of crystal violet-stained cells and represent the mean of independent experiments ± SD (n = 3). (C) Migration assay performed in Boyden chambers as described in the legend of Fig. 2E. The results are expressed as the means ± SD of three independent experiments performed in triplicate. (D) HUVEC as described in panel A were wounded with a sterile pipette tip, washed with culture medium, and incubated in complete medium on vitronectin or gelatin-coated plates as indicated. Cells were observed under a light microscope and photographed at 0 and 8 h. A representative experiment is shown (original magnification, ×50). (E) HUVEC as described in panel A were plated on BD Matrigel and incubated in complete medium for 20 h for in vitro angiogenesis assays. (F) Quantification of tube length was performed based on the results shown in panel E. Data are presented as means ± SD from four different fields randomly chosen from each group from three sets of experiments. (G) HUVEC as described in panel A were stained for phalloidin (green) and FAK (red). Scale bar, 30 μm.