Targeting strategy
Gene targeting strategy for B-hGPR20 mice. The exon 2 of mouse Gpr20 gene that encodes the full-length protein was replaced by human GPR20 exon 2 in B-hGPR20 mice.
mRNA expression analysis
Strain specific analysis of GPR20 mRNA expression in wild-type C57BL/6 mice and B-hGPR20 mice by RT-PCR. Colon RNA was isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hGPR20 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human GPR20 primers. Mouse Gpr20 mRNA was detectable only in wild-type C57BL/6 mice. Human GPR20 mRNA was detectable only in homozygous B-hGPR20 mice but not in wild-type mice.
Protein expression analysis
Strain specific GPR20 expression analysis in homozygous B-hGPR20 mice by western blot. Stomach, colon, bladder, lung, liver, spleen, and brain were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hGPR20 mice (H/H), and analyzed by western blot with anti-GPR20 antibody. m/hGPR20 was detectable in colon of wild-type C57BL/6 mice and homozygous B-hGPR20 mice.
