Gene targeting strategy for B-hADAMTS5 mice. The exons 1-8 of mouse Adamts5 gene that encode the full-length protein were replaced by human ADAMTS5 exons 1-8 in B-hADAMTS5 mice.

Western blot analysis of ADAMTS5 protein expression in homozygous B-hADAMTS5 mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hADAMTS5 mice (H/H) , and then analyzed by western blot with anti-ADAMTS5 antibody. 40 μg total proteins were loaded for western analysis. The antibody cross recognizes human and mouse ADAMTS5. ADAMTS5 was detected in kidney, testis, lung, spleen, and brain.

Strain specific analysis of ADAMTS5 mRNA expression in wild-type C57BL/6 mice and B-hADAMTS5 mice by RT-PCR. Testis RNA was isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hADAMTS5 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human ADAMTS5 primers. Mouse Adamts5 mRNA was detectable only in wild-type C57BL/6 mice. Human ADAMTS5 mRNA was detectable only in homozygous B-hADAMTS5 mice but not in wild-type mice.