Wound-healing migration assay

The assay was performed as previously described [5, 10]. HUVECs were seeded in precoated 6-well plates, cultured to 90%–100% and starved in EBM-2 medium with 0.2% serum for 3 h. Scratched lines were created using 200-μl pipette tips. Floating cells were removed with PBS. After incubation with PD98059 (10 μM) or mock control for 2 h in EBM-2 medium with reduced FBS, cells were further incubated with VEGF or Scg3 at indicated concentrations for additional 20 h. Images were taken under a light microscope at 0 and 20 h (before and after growth factor treatment). Wound closure was analyzed by counting cells migrated into the denuded region.