Figure 4
Figure 4. VWF-CFP type 2N variants induce formation of storage organelles that recruit both factor VIII-YFP and P-selectin. HEK293 cells stably expressing FVIII-YFP were transfected with P-selectin and wild-type VWF-CFP (A), P-selectin and VWF type 2N variants that display a moderately reduced binding to FVIII (Arg854Gln, Cys1060Arg) (B), or P-selectin and VWF type 2N variants that display a severely reduced binding to FVIII (Arg763Gly, Thr791Met, Arg816Trp) (C). Cells were stained for P-selectin using rabbit polyclonal anti–human CD62P antibody. Shown is triple fluorescent detection of representative 3-dimensional projections (Z-stacks 0.4 μm). VWF-CFP, FVIII-YFP, and P-selectin are shown in red, green, and blue, respectively. Triple fluorescent detection is shown in the color merges (regions of triple colocalization are shown in white). The scale bar represents 10 μm.

VWF-CFP type 2N variants induce formation of storage organelles that recruit both factor VIII-YFP and P-selectin. HEK293 cells stably expressing FVIII-YFP were transfected with P-selectin and wild-type VWF-CFP (A), P-selectin and VWF type 2N variants that display a moderately reduced binding to FVIII (Arg854Gln, Cys1060Arg) (B), or P-selectin and VWF type 2N variants that display a severely reduced binding to FVIII (Arg763Gly, Thr791Met, Arg816Trp) (C). Cells were stained for P-selectin using rabbit polyclonal anti–human CD62P antibody. Shown is triple fluorescent detection of representative 3-dimensional projections (Z-stacks 0.4 μm). VWF-CFP, FVIII-YFP, and P-selectin are shown in red, green, and blue, respectively. Triple fluorescent detection is shown in the color merges (regions of triple colocalization are shown in white). The scale bar represents 10 μm.

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