Basic Information
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Targeting strategy
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Gene targeting strategy for B-hLGR4 mice. The exons 5~18 of mouse Lgr4 gene were replaced by human LGR4 exons 5~18 in B-hLGR4 mice.
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Protein expression analysis
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Western blot analysis of LGR4 protein expression in homozygous B-hLGR4 mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hLGR4 mice (H/H), and then analyzed by western blot with anti-LGR4 antibody. 60 μg total proteins were loaded for western blotting analysis. LGR4 protein was detected in the lung, testis, colon, spleen, kidney of wild-type C57BL/6 mice and homozygous B-hLGR4 mice, as the antibody is cross-recognize both human and mouse LGR4.
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mRNA expression analysis
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Strain specific analysis of LGR4 mRNA expression in wild-type C57BL/6 mice and B-hLGR4 mice by RT-PCR. Kidney RNA were isolated from wildtype C57BL/6 mice (+/+) and homozygous B-hLGR4 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human LGR4 primers. Mouse LGR4 mRNA was detected in wild-type C57BL/6 mice. Human LGR4 mRNA was detected in homozygous B-hLGR4 mice but not in wild-type mice.
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Summary
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mRNA expression analysis:
Mouse Lgr4 mRNA was detected in kidney of wild-type C57BL/6 mice. Human LGR4 mRNA was detected in kidney of B-hLGR4 mice.
Protein expression analysis:
LGR4 protein was detected in the lung, testis, colon, spleen, kidney of wild-type C57BL/6 mice and homozygous B-hLGR4 mice, as the antibody is cross-recognize both human and mouse LGR4.
