Deposition of plasma fibronectin on platelets adherent to collagen or VWF under static conditions. (A) Coverslips were coated with fibrillar type I collagen (CL), VWF, both proteins simultaneously, or collagen and then VWF. Platelets and 25 μg/mL (50 nM) FITC-fibronectin (FITC-FN) without or with 1 μM FUD and without LPA were added and incubated for 1 hour. Platelets were stained with rhodamine-phalloidin and then examined by epifluorescent microscopy for distribution of FITC-fibronectin and filamentous actin cytoskeleton. Platelets bound to collagen as aggregates. The images were obtained at the level of the coverslip surface. Bar = 10 μm. (B) Platelets were plated on protein-coated wells of a 96-well plate and incubated with or without 50 μg/mL soluble fibronectin (pFN) for 30 minutes without LPA. After 3 rinses, adherent platelets were quantified as described in “Materials and methods.” Values represent the mean ± SD (n = 3).