CLEC-2 deficiency impairs platelet aggregation on immobilized VWF under flow. Washed platelets treated with albumin or 20 μg/mL of RGD, an integrin αIIbβ3 antagonist, were perfused over immobilized plasma VWF (150 μg/mL) at 10 dyne/cm2 for 10 minutes. After removing nonadherent platelets, adhered platelets were fixed with 2% paraformaldehyde (PFA), stained with anti-CD41 antibodies, and observed with microscopy. (A) Representative images of platelet aggregation on immobilized VWF under flow. (B) Quantification of platelet aggregation on immobilized VWF under flow. Adherent platelets described in (A) were quantified by analyzing the pixels of CD41-positive areas in each field. The data are representative of 5 independent experiments, and data represent mean ± SD. *P < .05.
