Figure 6.
Membrane protein recruitment by WT and mutant rVWF. HEK293 cells were transiently cotransfected with either WT rVWF (A,E); rVWF-Cys1225Gly (B,F); rVWF-Cys788Arg (C,G); or rVWF-Arg273Trp (D,H). The cells were also transiently cotransfected with EGFP-CD63 (A-D) or HRP-P-selectin (E-H), and were immunolabeled for VWF (in red, panels A-H) and HRP (in green, panels E-H) 2 days later. The GFP in panels A-D (in green) was visible without staining. In each panel, the majority of pseudo-WPBs are yellow (arrowheads), indicating a colocalization of VWF and the marker. CD63 and P-selectin are also visible in other vesicles (as described in “Membrane protein recruitment”), where there is no colocalization with VWF. Scale bar measures 20 μm.