Figure 2. Effect of platelet inhibitors on platelet adhesion and spreading on nidogen-1.

Human washed platelets (2 × 10⁷/mL) were pre-treated with A. vehicle (DMSO), Syk inhibitor (Bay-61–3606, 5 μM), SFK inhibitor (PP2, 10 μM), PI3K inhibitor (wortmannin, 100 nM), C. PLC inhibitor (U73122, 10 μM), PLC inhibitor analog (U73343,10 μM) or IKK inhibitor (IKK-16, 10 μM) E. vehicle (DMSO) or anti-GPVI (ACT017 GPVI inhibitor, 40 μg/mL) H. vehicle (DMSO), anti- β₁ (AIIB2, 20 μg/mL), anti-αIIbβ3 (integrillin, 20 μg/mL) (data not shown) or ADP scavenger (apyrase, 2U/ml) for 15 min prior to seeding on coverslips coated with recombinant nidogen-1 (100 μg/mL) for 45 min at 37° C and imaged using differential interference contrast (DIC) microscopy. Images are representative of at least 3 independent experiments. B and D. Number of adherent platelets and mean surface area of platelet on nidogen-1 were recorded for 3 fields of view for each condition and expressed as mean ± SEM. F and H. Number of adherent platelets was recorded for 3 fields of view for each condition and expressed as mean ± SEM. * P < 0.05 with respect to platelet adhesion in the absence of inhibitors. Scale bar, 10 μm.