Strain specific analysis of LGR5 mRNA expression in wild-type C57BL/6 mice and B-hRSPO3hLGR5 mice by RT-PCR. Stomach RNA was isolated from wildtype C57BL/6 mice (+/+) and homozygous B-hRSPO3hLGR5 mice (H/H; H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human LGR5 primers. Mouse Lgr5 mRNA was detectable only in wild-type C57BL/6 mice. Human LGR5 mRNA was detectable only in homozygous B-hRSPO3hLGR5 mice but not in wild-type mice. 

Strain specific analysis of RSPO3 mRNA expression in wild-type C57BL/6 mice and B-hRSPO3hLGR5 mice by RT-PCR. Stomach RNA was isolated from wildtype C57BL/6 mice (+/+) and homozygous B-hRSPO3hLGR5 mice (H/H; H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human RSPO3 primers. Mouse Rspo3 mRNA was detectable only in wild-type C57BL/6 mice. Human RSPO3 mRNA was detectable only in homozygous B-hRSPO3hLGR5 mice but not in wild-type mice. 

Western blot analysis of LGR5 protein expression in homozygous B-hRSPO3hLGR5 mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hRSPO3hLGR5 mice (H/H; H/H), and then analyzed by western blot with anti-LGR5 antibody. 40 μg total proteins were loaded for western blotting analysis. LGR5 was detected in stomach, colon, frontal lobe, kidney, liver and lung.

Western blot analysis of RSPO3 protein expression in homozygous B-hRSPO3hLGR5 mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hRSPO3hLGR5 mice (H/H; H/H), and then analyzed by western blot with anti-RSPO3 antibody. 40 μg total proteins were loaded for western blotting analysis. RSPO3 was detected in stomach.