Effect of WT and mutated mVWF on bleeding time in VWF^−/− mice after hydrodynamic gene delivery. VWF^−/− mice were injected with 50 μg of WT pLIVE-mVwf or pLIVE-mVwf mutated in the RGD sequence (D2509G), in the collagen-binding site (D1742A, S1783A, H1786A), or in the GPIb-binding site (K1362A). Bleeding time was assessed 3 days after injection by tail tip transection. Observation was stopped at 600 seconds when bleeding did not cease. Each symbol represents one mouse, and the median value of each group is represented by a horizontal line. pLIVE stands for mice injected with the vector only. NS indicates not significant.