VWF binding to macrophages is concentration-dependently inhibited by BT200. (A) Binding of full-length recombinant human VWF to murine BMDMs was assessed by flow cytometry in the presence or absence of increasing concentrations of BT200. Magenta is VWF binding in the absence of BT200; gray is background in the absence of VWF; blue to red signals for increasing BT200 concentrations. The y-axis represents the binding capacity normalized to the number of cells. FITC-A on the x-axis represents the fluorescence intensity absorbance at the wavelength of 490 nm, in which higher values indicate more VWF binding. (B-E) BMDM or primary human monocyte–derived macrophage binding on the y-axis are calculated from mean fluorescent intensities at different BT200 or BT100 concentrations presented on the x-axis. Error bars depict the standard deviation of the fluorescent intensities. P values are outcomes of extra-sum-of-squares F test and represent whether the binding curves demonstrate a significant binding to BMDMs in panels B-D or to compare the binding capacity of BT100 and BT200 to BMDMs in panel E. (B) Full-length human rVWF binding (%) to BMDMs is concentration-dependently inhibited by BT200. (C) Full-length human rVWF binding (%) to primary human macrophages is concentration-dependently inhibited by BT200. (D) Binding of truncated human VWF-A1A2A3 to BMDMs is concentration-dependently inhibited by BT200. (E) Binding of full-length human rVWF to BMDMs is significantly more inhibited by the pegylated BT200 compared to the unpegylated BT100. FITC-A, fluorescein isothiocyanate; rVWF, recombinant VWF.