Stimulation of endothelial cells with Ang1 induced SMC migration through HGF up-regulation. (A) Coculture assay using Transwell was developed to evaluate SMC migration. Both sides of the filters were coated with Matrigel. ECs were seeded underneath the filter, and SMCs were seeded in the upper chamber. (B) HUVECs were infected with various viral vectors expressing genes of interest for 48 hours, and SMCs were labeled with RFP. Then the coculture assay was set up as shown in panel A. Migrated SMCs on the other side of the filter were counted through a microscope in randomly selected high-power fields. (C) Effects of neutralization of HGF function in EC-SMC recruitment were evaluated with the coculture assay. Neutralizing HGF antibody or control IgG was added 1 hour before HASMCs were added to the wells. Experiments were performed at least 3 times in more than 2 wells for each treatment. Five fields were counted for each filter in each experiment. *P < .01 compared with control.