Basic Information
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Targeting strategy
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Gene targeting strategy for B-hLRP5 mice.
The exons 2~20 of mouse Lrp5 gene that encode the extracellular domain were replaced by human LRP5 exons 2~20 in B-hLRP5 mice. -
Protein expression analysis
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Western blot analysis of LRP5 protein expression in homozygous B-hLRP5 mice. Various tissue lysates were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hLRP5 mice (H/H), and then analyzed by western blotting with anti-LRP5 antibody. 40μg total proteins were loaded for western blotting analysis. LRP5 protein was detected in the liver, lung and kidney of wild-type C57BL/6JNifdc mice and homozygous B-hLRP5 mice, as the antibody showed cross-recognition both for human and mouse LRP5.
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mRNA expression analysis
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Strain specific analysis of LRP5 mRNA expression in wild-type C57BL/6JNifdc mice and B-hLRP5 mice by RT-PCR. Lung RNA were isolated from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hLRP5 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human LRP5 primers. Mouse Lrp5 mRNA was detectable only in wild-type C57BL/6JNifdc mice. Human LRP5 mRNA was detectable only in homozygous B-hLRP5 mice but not in wild-type mice.
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Summary
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mRNA expression analysis:
Mouse Lrp5 mRNA was detectable in lung of wild-type C57BL/6JNifdc mice. Human LRP5 mRNA was detectable in B-hLRP5 mice, and was confirmed via Sanger Sequencing.Protein expression analysis:
Human and mouse LRP5 was both detectable on lung, kidney and liver of WT mice and homozygous B-hLRP5 mice, as the antibody showed cross-recognition.
