Basic Information
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Targeting strategy
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Gene targeting strategy for B-hEPHB3 mice.
The exons 3~7 of mouse Ephb3 gene that encode the extracellular domain were replaced by human EPHB3 exons 3~7 in B-hEPHB3 mice.
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Protein expression analysis
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Western blot analysis of EPHB3 protein expression in homozygous B-hEPHB3 mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hEPHB3 mice (H/H), and then analyzed by proteinsimple-western blot with anti-EPHB3 antibody. 3μg total proteins were loaded for western blotting analysis. The EPHB3 protein was detected in the lung, liver, stomach, brain of wild-type C57BL/6 mice and homozygous B-hEPHB3 mice, as confirmed by cross-reactivity of the antibody against mouse and human EPHB3 proteins.
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mRNA expression analysis
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Strain specific analysis of EPHB3 mRNA expression in wild-type C57BL/6 mice and B-hEPHB3 mice by RT-PCR. Stomach RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hEPHB3 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human EPHB3 primers. Mouse Ephb3 mRNA was detectable only in wild-type C57BL/6 mice. Human EPHB3 mRNA was detectable only in homozygous B-hEPHB3 mice but not in wild-type mice.
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Summary
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mRNA expression analysis:
Mouse Ephb3 mRNA was detectable in stomach of wild-type C57BL/6 mice. Human EPHB3 mRNA was detectable in B-hEPHB3 mice.
Protein expression analysis:
EPHB3 protein was detected in the lung, liver, stomach, brain of wild-type C57BL/6 mice and homozygous B-hEPHB3 mice, as confirmed by cross-reactivity of the antibody against mouse and human EPHB3 proteins.