Gene targeting strategy for B-hPTHrP mice. The exons 2-4 of mouse Pthrp gene that encode the full-length protein were replaced by human PTHRP exons 4-6 in B-hPTHrP mice.

Western blot analysis of PTHrP protein expression in homozygous B-hPTHrP mice. Various tissue lysates were collected from C57BL/6 wildtype (+/+) mice and homozygous B-hPTHrP (H/H) mice, and then analyzed by western blot with anti-PTHrP antibody. 40 μg total proteins were loaded for western blotting analysis. PTHrP was detected in brain, stomach, kidney and spleen.

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