Recognition of uPA domains by α_Mβ₂ and uPAR.

HEK293 cell lines (A) and unstimulated or PMA-stimulated (20 nM) PMNs (B) were allowed to adhere to microtiter plates coated with HMW-tc-uPA or its domains for 30 minutes at 37°C. Nonadherent cells were removed by washing with PBS, and the number of adherent cells was quantitated as described in “Materials and methods.” HEK293 cells expressing uPAR or α_Mβ₂ or both (C) and PMA-stimulated PMNs (D) were added to the transwells (2 × 10⁵cells/well) and allowed to migrate for 6 hours at 37°C in 5% CO₂. HMW-tc-uPA or its domains were added to the lower chamber of the transwell at 100 nM (C) and 10 nM (D) concentrations. The data are means ± SEM of triple measurements from 3 independent experiments.