Basic Information
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Targeting strategy
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Gene targeting strategy for B-hTEK/hTIE1 mice.
Exons 2~14 of the mouse Tie1 gene that encode the full-length protein were replaced by human TIE1 exons 2~14 in B-hTEK/hTIE1 mice.
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Protein expression analysis-TEK
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Strain specific TEK expression analysis in homozygous B-hTEK/hTIE1 mice by flow cytometry. Lung were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hTEK/hTIE1 mice (H/H, H/H), and analyzed by flow cytometry with anti-TEK antibody. mTEK was detectable in wild-type, hTEK was detectable in wild-type and homozygous B-hTEK/hTIE1 mice, as the antibody is cross-recognize both human and mouse TEK.
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Protein expression analysis-TIE1
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Strain specific TIE1 expression analysis in homozygous B-hTEK/hTIE1 mice by flow cytometry. Lung were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hTEK/hTIE1 mice (H/H, H/H), and analyzed by flow cytometry with anti-TIE1 antibody. hTIE1 was detectable in wild-type and homozygous B-hTEK/hTIE1 mice, as the antibody is cross-recognize both mouse and human TIE1.
Western blot analysis of TIE1 protein expression in homozygous B-hTEK/hTIE1 mice. Various tissue lysates were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hTEK/hTIE1 mice (H/H), and then analyzed by western blot with anti-TIE1 antibody. 40 μg total proteins were loaded for western blotting analysis. TIE1 was detected in lung, kidney, spleen and heart in wild-type mice and homozygous B-hTEK/hTIE1 mice, as the antibody is cross-recognize both human and mouse TIE1.
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mRNA expression analysis
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Strain specific analysis of TEK mRNA expression in wild-type C57BL/6 mice and B-hTEK/hTIE1 mice by RT-PCR. Lung RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hTEK/hTIE1 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human TEK primers. Mouse Tek mRNA was detectable only in wild-type C57BL/6 mice. Human TEK mRNA was detectable only in homozygous B-hTEK/hTIE1 mice but not in wild-type mice.
Strain specific analysis of TIE1 mRNA expression in wild-type C57BL/6 mice and B-hTEK/hTIE1 mice by RT-PCR. Lung RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hTEK/hTIE1 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human TIE1 primers. Mouse Tie1 mRNA was detectable only in wild-type C57BL/6 mice. Human TIE1 mRNA was detectable only in homozygous B-hTEK/hTIE1 mice but not in wild-type mice.
