PMC

E17.5 FV(λ/λ) embryos with PZ(−/−), PZ(+/−), and PZ(+/+) genotypes.

Antifibrin(ogen) staining of tissues from E17.5 embryos. (Upper) Thrombosis (arrows) of spinal (Left), chest wall (Center), and pulmonary (Right) vessels in a FV(λ/λ)/PZ(−/−) embryo. (Lower) Hepatic fibrin deposition in FV(λ/λ) embryos with PZ(−/−) (Left), PZ(+/−) (Center), and PZ(+/+) (Right) genotypes.

Regulation of coagulation by PZ. (A) Factor Xa activity. Factor IXa (0.5 nM) was used to induce coagulation in mixtures containing rabbit brain phospholipids (cephalin, 15 μM); CaCl₂ (10 mM); barium-adsorbed, oxalated, pooled normal plasma (50% vol/vol); and factor X (100 nM) with or without PZ (30 nM). Phospholipids and CaCl₂ with or without PZ were incubated 2 min before the addition of the remaining reagents, and the reaction was initiated by the addition of the plasma. (B) Thrombin generation. Reactions were the same as described for A, except prothrombin (700 nM) was also added to each mixture. ■, −PZ; ●, +PZ.

Generation of PZ(−/−) mice. (A) Gene targeting. The targeting construct (Top) contains a neo cassette (neo) that replaces a 139-bp portion of exon 2 and induces a frameshift mutation. A herpes simplex virus–thymidine kinase cassette (tk) was added at the 3′ end of the mouse PZ DNA to permit negative selection. The predicted product of homologous recombination is shown (Bottom). The position of the ≈500-bp hybridization probe used to detect successful gene targeting is also depicted. (B) Southern blot analysis. Genomic DNA prepared from tail biopsies was analyzed by restriction digestion with XbaI and hybridization with the probe. Expected DNA fragment sizes are 9.0 kb for the wild-type allele and 3.2 kb for the disrupted allele. (C) Western blot analysis of mouse plasma with rabbit anti-human PZ polyclonal antibodies.