Effect of FVIIa and other agonists on cancer cell migration: involvement of PAR-2 and IL-8 in FVIIa-induced cell migration. MDA-MB-231 cells were placed in the upper well, and various concentrations of FVIIa (A) or other agonists (B) or FVIIa and antibodies against IL-8 or PAR-2 (C) were added to the lower well. In additional experiments, ASIS or anti-TF IgG was included with FVIIa in the lower well (A). The number of cells that migrated to the underside of the membrane in 20 hours at 37°C was determined as described in “Materials and methods” (mean ± SEM, n = 3 to 5). Concentrations of various reagents were: FVIIa, 50 nM (unless specified otherwise); ASIS, 500 nM; anti-TF IgG, 100 μg/mL; thrombin, 10 nM; trypsin, 10 nM; plasmin, 50 nM; PAR-1 AP, 50 μM; PAR-2 AP, 50 μM; rabbit antihuman IL-8 IgG, 50 μg/mL; rabbit anti–PAR-2 IgG, 500 μg/mL; control IgG, 50 μg/mL (1) or 500 μg/mL (2); and IL-8, 100 ng/mL. ^*Difference in value is statistically significant (P < .05) from the value obtained in control treatment (panel B) or corresponding control IgG (panel C).