Effects of 14-kDa ΔN-rhChM-I on the VEGF-A-induced chemotactic migration of HUVECs.

<p>Biological activity of G-rhChM-I and ΔN-rhChM-I was assessed by a modified Boyden chamber assay <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0094239#pone.0094239-Miura1" target="_blank">[8]</a>. Serum-starved HUVECs (7×10⁴ cells) were preincubated for 30 min with G-rhChM-I (•) or ΔN-rhChM-I (▴), and seeded onto vitronectin-coated cell culture inserts in serum-free medium. Chemotactic migration of HUVECs was induced by the addition of VEGF-A (20 ng/ml) in the lower chamber, and the cells were allowed to migrate for 4 h toward VEGF-A. The number of cells that had migrated to the bottom surface of the insert was counted. Values are the percentages of migrated cell numbers in the control culture that was stimulated by VEGF-A, and are the means ± SD of a triplicate assay. The data are representative of three independent experiments with similar results.