Basic Information
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Targeting strategy
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Gene targeting strategy for B-hCLDN3/hCLDN4 mice.
Mouse Cldn3 and Cldn4 gene were replaced by human CLDN3 and CLDN4 gene in B-hCLDN3/hCLDN4 mice.
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Protein expression analysis
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Western blot analysis of CLDN3 and CLDN4 protein expression in homozygous B-hCLDN3/hCLDN4 mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCLDN3/hCLDN4 mice (H/H, H/H), and then analyzed by western blot with cross-reactive anti-CLDN3 antibody or species-specific anti-CLDN4 antibody. 40 μg total proteins were loaded for western blotting analysis. CLDN3 and CLDN4 were detected in lung and colon.
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mRNA expression analysis
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Strain specific analysis of CLDN3 and CLDN4 mRNA expression in wild-type C57BL/6 mice and B-hCLDN3/hCLDN4 mice by RT-PCR. Colon RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hCLDN3/hCLDN4 mice (H/H, H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human CLDN3 or CLDN4 primers. Mouse Cldn3 and Cldn4 mRNA were detectable only in wild-type C57BL/6 mice. Human CLDN3 and CLDN4 mRNA were detectable only in homozygous B-hCLDN3/hCLDN4 mice but not in wild-type mice.
