NIF inhibits PMN adhesion to VWF and its derivatives. Freshly isolated PMNs were stimulated with PMA (100 nM) for 15 minutes, followed by an additional 15-minute incubation in the absence or presence of NIF (10 μg/mL). (A) Static adhesion of PMNs to immobilized VWF, VWF/D′-D3, or VWF/A1-A2-A3 was performed and quantified as described in the Figure 1 legend. Relative adhesion is plotted with adhesion to VWF as a reference (100%) in each panel. Control represents adhesion to BSA-coated wells. Data represent mean ± SEM of 4 experiments performed in duplicate. (B) Perfusion of PMA-stimulated PMNs to immobilized VWF was performed and analyzed as described in the Figure 2B legend. Cumulative number of adherent cells was plotted in the absence (○) or presence (•) of NIF (10 μg/mL). Data represent mean ± SEM of 4 experiments and are corrected for adhesion to PVP-coated coverslips (less than 2 cells/mm2 after 6 minutes).
