Figure 5.

AVA4746 attenuates tube formation by HUVECs. HUVECs treated with either 0, 5 or 25 µM AVA4746 for 30 min were seeded into Matrigel-coated plates for 4-6 h. (A) Representative flow cytometry dot plot of human integrin α4 and β1 expression in HUVECs before seeding into the Matriel wells. (B) Representative images of the HUVECs are shown. (C) Tube formation was assessed by analyzing the number of nodes (pixels with ≥ three neighboring elements corresponding to a bifurcation), segments (elements delimited by two junctions), meshes (areas enclosed by segments or master segments and made by tube-like structures) and total area, before being subsequently quantified. Images were analyzed with using the ImageJ software and the ‘angiogenesis analyzer’ plugin. ^*P<0.05, ^**P<0.01 and ^***P<0.001. HUVEC, Human umbilical vein endothelial cells; α4, α4 intergrin; β1, β1 integrin; Nb, number.