Basic Information
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Targeting strategy
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Gene targeting strategy for B-hRSPO3 mice.
The exons 2~5 of mouse Rspo3 gene were replaced by human RSPO3 exons 2~5 in B-hRSPO3 mice.
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Protein expression analysis
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Western blot analysis of RSPO3 protein expression in homozygous B-hRSPO3 mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hRSPO3 mice (H/H), and then analyzed by western blot with anti-RSPO3 antibody. 60 μg total proteins were loaded for western blotting analysis. RSPO3 protein was detected in the stomach of wild-type C57BL/6 mice and homozygous B-hRSPO3 mice, as the antibody is cross-recognize both human and mouse RSPO3.
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mRNA expression analysis
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Strain specific analysis of RSPO3 mRNA expression in wild-type C57BL/6 mice and B-hRSPO3 mice by RT-PCR. Stomach RNA were isolated from wildtype C57BL/6 mice (+/+) and homozygous B-hRSPO3 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human RSPO3 primers. Mouse Rspo3 mRNA was detected in wild-type C57BL/6 mice. Human RSPO3 mRNA was detected in homozygous B-hRSPO3 mice but not in wild-type mice.
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Summary
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mRNA expression analysis:
Mouse Rspo3 mRNA was detected in stomach of wild-type C57BL/6 mice. Human RSPO3 mRNA was detected in B-hRSPO3 mice.
Protein expression analysis:
RSPO3 protein was detected in the stomach of wild-type C57BL/6 mice and homozygous B-hRSPO3 mice, as the antibody is cross-recognize both human and mouse RSPO3.
