Gene targeting strategy for B-hTGR5 mice. The exons 1-2 of mouse Tgr5 gene that encode the 5’UTR, promoter, signal peptide, extracellular domain, transmembrane domain, cytoplasmic region and 3’UTR are replaced by human counterparts in B-hTGR5 mice. The TGR5 expression is driven by endogenous human TGR5 promoter, while mouse Tgr5 gene transcription and translation will be disrupted. 

Western blot analysis of TGR5 protein expression in homozygous B-hTGR5 mice. Various tissue lysates were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hTGR5 mice (H/H), and then analyzed by western blot with anti-TGR5 antibody (abcam, ab72608). 40 μg total proteins were loaded for western blotting analysis. TGR5 was detected in Lung, spleen, bladder, brain.

Strain specific analysis of TGR5 mRNA expression in wild-type C57BL/6JNifdc mice and B-hTGR5 mice by RT-PCR. Lung RNA were isolated from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hTGR5 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human TGR5 primers. Mouse Tgr5 mRNA was only detectable in wild-type mice. Human TGR5 mRNA was exclusively detectable in homozygous B-hTGR5 mice but not in wild-type mice.