Gene targeting strategy for B-hSUCNR1 mice. The exons 1~2 of mouse Sucnr1 gene that encode full-length protein were replaced by human SUCNR1 exons 2~3 in B-hSUCNR1 mice.

Strain specific analysis of SUCNR1 mRNA expression in wild-type C57BL/6N mice and B-hSUCNR1 mice by RT-PCR. Liver RNA were isolated from wild-type C57BL/6N mice (+/+) and homozygous B-hSUCNR1 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human SUCNR1 primers. Mouse Sucnr1 mRNA was detectable only in wild-type C57BL/6N mice. Human SUCNR1 mRNA was detectable only in homozygous B-hSUCNR1 mice but not in wild-type mice.

Western blot analysis of SUCNR1 protein expression in homozygous B-hSUCNR1 mice. Various tissue lysates were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hSUCNR1 mice (H/H), and then analyzed by western blot with anti-SUCNR1 antibody. 4 μg total proteins were loaded for western blotting analysis. SUCNR1 protein was detected in the spleen，liver and kidney of wild-type C57BL/6N mice and homozygous B-SUCNR1 mice, as the antibody is cross-recognize both human and mouse SUCNR1.