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. 2009 Sep 16;284(46):31704–31713. doi: 10.1074/jbc.M109.030312

FIGURE 3.

FIGURE 3.

Complex FVIII·vWF can bind to RANKL and OPG prevents its binding. A, in the pH 7.4 buffer, FVIII·vWF complex, but not recombinant Factor VIII, binds to RANKL. FVIII·vWF complex (50 units/ml) or recombinant Factor VIII (50 units/ml) was injected at a flow rate of 10 μl/min over the immobilized RANKL sensor chip for 5 min, and the dissociation was monitored for 10 min. B, shown is RANKL (500, 100, and 10 ng/ml) bound to the coated FVIII·vWF complex (1 unit/ml) using an ELISA assay. Results are expressed in arbitrary units. C, FVIII·vWF complex increases the binding of OPG to RANKL. OPG was incubated with increasing concentrations of FVIII·vWF complex for 2 h before the injection to immobilized RANKL. D, FVIII·vWF complex, OPG, and RANKL can form a tripartite complex. Human RANKL (5 μg/ml) was injected to immobilized OPG with a flow rate of 20 μl/min, then the FVIII·vWF complex was injected. Schematic explanations are represented in c and d.