Fig. 9. Hep I modulates aFGF/bFGF-induced endothelial cell chemotaxis. BAE cells were plated at low density under serum-free conditions onto glass coverslips coated with vitronectin and fibronectin. Cells were allowed to attach for 3 hours. Some coverslips were pretreated for 30 minutes with 100 nM hep I or 7.8 nM TSP1. Coverslips were loaded onto the Dunn Chamber in serum-free media (A,D,G) or serum-free media containing either 100 nM hep I (B,E,H) or 7.8 nM TSP1 (C,F,I). Media was removed from the outer well and corresponding media containing aFGF (67 pM) (D,E,F; Movies 4 and 6, available at jcs.biologists.org/supplemental) or bFGF (61 pM) (G,H,I; Movies 5 and 7, available at jcs.biologists.org/supplemental) was then loaded into the outer well establishing a chemical gradient. The chamber was sealed, and time-lapse video was taken of the cells over a 7-hour time span. Migration of individual cells was tracked using Metamorph software and individual tracks were analyzed for distance, displacement, directionality and orientation. Final positions of the cells are plotted with the initial point being the origin and the top of the graph representing the outer well. Results are representative of at least six separate experiments. Average displacement towards the outer well (X) is given for each treatment. *P<0.05, **P<0.01.