Gene targeting strategy for B-hVWF mice. The part of exon 2 and all of exons 3 of mouse Vwf gene sequences were replaced by human VWF coding sequences that encodes the full-length protein, 3’UTR of mouse Vwf gene and STOP termination sequences in B-hVWF mice. The insertion disrupts the endogenous murine Vwf gene, resulting in a non-functional transcript.

Strain specific analysis of VWF gene expression in wild type (WT) mice and B-hVWF mice by RT-PCR. Mouse Vwf mRNA was detectable only in lung of WT mice (+/+). Human VWF mRNA was detectable only in homozygous B-hVWF mice (H/H) but not in WT mice (+/+).

Strain specific VWF expression analysis in wild type (WT) mice and B-hVWF mice by ELISA. Plasma were collected from WT mice (+/+) and homozygous B-hVWF mice (H/H), and analyzed by ELISA with VWF ELISA kit. Mouse VWF was detectable in WT mice (+/+) and homozygous B-hVWF mice (H/H) due to the cross-reactivity of antibodies. Human VWF was exclusively detectable in homozygous B-hVWF mice (H/H) but not in WT mice (+/+) . Values are expressed as mean ± SEM. ND: not detectable.