Adhesion of VWF-coated beads to immobilized LRP1 under flow. Fluoresbrite-YG microspheres were coated with BSA or VWF (25, 50, or 200 μg/1.4 × 109 beads) and perfused over BSA- or LRP1-coated channels (shear force: 2.5, 5, or 10 dyne/cm2) using Bioflux microfluidic equipment. (A) Snapshots of VWF- or BSA-coated microspheres being perfused (t = 2 minutes) over LRP1- or BSA-coated channels. (B) Quantification of adhesion of beads coated with 50 μg/1.4 × 109 beads at shear forces of 2.5, 5, or 10 dyne/cm2. (C) Quantification of adhesion of beads coated with 25, 50, or 200 μg/1.4 × 109 beads at 5 dyne/cm2. Inset, Quantification of adhesion of beads coated with 50 μg/1.4 × 109 beads at 5 dyne/cm2 in the absence or presence of RAP (20 μg/mL). Adhesion was quantified by counting the number of adhered microspheres in 5 microscopic fields taken after 2.5 minutes of perfusion within a period of 15 seconds using ImageJ 1.44 software. Data represent the mean ± SEM of 3 to 6 perfusions. Closed circles indicate VWF beads and LRP1 coating; open circles, VWF beads and BSA coating; closed squares, BSA beads and LRP1 coating; and open squares, BSA beads and BSA coating.
