PMN adhesion and migration to the TSP-4 variants. (A) The 96-well plates were coated with increasing concentrations of TSP-4 variants for 16 hours at 4°C. Resting or PMA-stimulated (1 nM) PMNs were allowed to adhere for 30 minutes at 37°C. Adherent cells were detected using the Cyquant Cell Proliferation kit. (B) The lower filter surface of Boyden chambers was coated with increasing concentrations of TSP-4 variants. Resting or PMA-stimulated PMNs were added to the upper chamber and incubated for 6 hours at 37°C and 5% CO₂. The migrated cells were counted using the DNA-based Cyquant Cell Proliferation kit. The data are means ± SEMs of triple measurements from 3 blood donors.