Figure 1. Adhesion and spreading of human platelets on nidogen-1.

A. Human washed platelets (2 × 10⁷/mL) were placed on coverslips coated with BSA (5 mg/mL), fibrinogen (100 μg/mL), soluble collagen (50 μg/mL), or recombinant human nidogen-1 (100 μg/mL) for 45 min at 37° C and imaged using differential interference contrast (DIC) microscopy. Images are representative of 3 independent experiments. Scale bar, 10 μm. B. The number of adherent platelets on BSA, fibrinogen (FG), soluble collagen (CL) and nidogen-1 (ND) for increasing platelet concentrations and increasing protein concentrations were recorded for 3 fields of view and expressed as mean ± SEM from at least 3 different experiments. C. Lysates from washed human platelets seeded on coverslips coated with recombinant nidogen-1, fibrinogen, CRP-XL (collagen related peptide), fibrillar collagen, or quiescent platelets (resting) in solution were analyzed for total phosphoprotein content with 4G10 or phosphorylation of Syk pY525, BTK pY223 (Bruton’s tyrosine kinase), BTK pY551, activation of PKC (protein kinase C), PLCγ2 pY1217 (phospholipase Cγ2), or activation of NF-κB through the analysis of IKK and p65 phosphorylation by western blotting (WB). Protein molecular markers on the right.