Targeting strategy
Gene targeting strategy for B-hFSHB mice. The exons 2-3 of mouse Fshb gene that encode the full-length protein were replaced by human FSHB exons 2-3 in B-hFSHB mice.
Protein expression analysis
Western blot analysis of FSHB protein expression in homozygous B-hFSHB mice. Various tissue lysates were collected from C57BL/6 wild-type mice (+/+) and homozygous B-hFSHB mice (H/H), and then analyzed by western blot with anti-FSHB antibody. 30 μg total proteins were loaded for western blotting analysis. Human FSHB was detected in pituitary gland of homozygous B-hFSHB mice.
mRNA expression analysis
Strain specific analysis of FSHB mRNA expression in wild-type C57BL/6 mice and homozygous B-hFSHB mice by RT-PCR. Pituitary gland RNA was isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hFSHB mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human FSHB primers. Mouse Fshb mRNA was detectable only in wild-type C57BL/6 mice. Human FSHB mRNA was detectable only in homozygous B-hFSHB mice but not in wild-type mice.
