Gene targeting strategy
Gene targeting strategy. The full coding region of mouse Plau was replaced by human PLAU coding region sequences in B-hPLAU/hPLAUR mice. The coding region except signal peptide of mouse Plaur gene was replaced by human PLAUR counterpart gene sequences in B-hPLAU/hPLAUR mice.
Protein expression analysis
Species-specific PLAU protein expression analysis in homozygous B-hPLAU/hPLAUR mice by ELISA. Serum was collected from wild-type C57BL/6 mice and homozygous B-hPLAU/hPLAUR mice (male, 9-week-old, n=3), and analyzed by ELISA with species-specific PLAU ELISA kits. Mouse PLAU were detectable in wild-type C57BL/6 mice. Human PLAU was exclusively detectable in homozygous B-hPLAU/hPLAUR mice but not in wild-type mice.
Species-specific PLAUR protein expression analysis in homozygous B-hPLAU/hPLAUR mice by flow cytometry. Bone marrow cells were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hPLAU/hPLAUR mice (H/H), and analyzed by flow cytometry with species-specific anti-PLAUR antibodies. Mouse PLAUR was only detected in dendritic cells from wild-type mice, while human PLAUR protein was detected in mono/macrophages, granulocytes, and dendritic cells from B-hPLAU/hPLAUR mice.