Basic Information
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Targeting strategy
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Gene targeting strategy for B-hEGFR/hMET mice.
The exons 2-17 of mouse Egfr gene that encode the extracellular domain were replaced by human EGFR exons 2-17 in B-hEGFR/hMET mice.
The exons 3-14 of mouse Met gene that encode the extracellular domain were replaced by human MET exons 2-13 in B-hEGFR/hMET mice. -
mRNA expression analysis
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Strain specific analysis of EGFR gene expression in wild type (WT) mice and B-hEGFR/hMET mice by RT-PCR. Mouse Egfr mRNA was detectable only in liver and kidney of WT mice (EGFR+/+MET+/+). Human EGFR mRNA was detectable only in B-hEGFR/hMET mice (EGFRH/HMETH/+) but not in WT mice (EGFR+/+MET+/+). The positive band was confirmed to be correct by sequencing.
Strain specific analysis of MET gene expression in wild type (WT) mice and B-hEGFR/hMET mice by RT-PCR. Mouse Met mRNA was detectable in liver and kidney of WT mice (EGFR+/+MET+/+) and B-hEGFR/hMET mice (EGFRH/HMETH/+). Human MET mRNA was detectable only in B-hEGFR/hMET mice (EGFRH/HMETH/+) but not in WT mice (EGFR+/+MET+/+). The positive band was confirmed to be correct by sequencing.
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Protein expression analysis
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Strain specific MET expression analysis in homozygous B-hEGFR/hMET mice by western blot. Liver and kidney were collected from wild type (WT) mice (EGFR+/+MET+/+) and B-hEGFR/hMET mice (EGFRH/HMETH/+), and analyzed by western blot with anti-MET antibody. MET was detectable in WT mice (EGFR+/+MET+/+) and B-hEGFR/hMET mice (EGFRH/HMETH/+) due to the cross-reactivity of antibodies.
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IHC analysis of EGFR expression
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Immunohistochemical (IHC) analysis of EGFR expression in homozygous B-hEGFR/hMET mice. The liver and kidney were collected from wild type (WT) mice (EGFR+/+MET+/+) and B-hEGFR/hMET mice (EGFRH/HMETH/+) and analyzed by IHC with anti-EGFR antibody. EGFR was detectable in WT mice (EGFR+/+MET+/+) and B-hEGFR/hMET mice (EGFRH/HMETH/+) due to the cross-reactivity of antibodies.
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Summary
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mRNA expression analysis:
Mouse Egfr mRNA was detectable only in liver and kidney of WT mice (EGFR+/+MET+/+). Human EGFR mRNA was detectable only in B-hEGFR/hMET mice (EGFRH/HMETH/+) but not in WT mice (EGFR+/+MET+/+). The positive band was confirmed to be correct by sequencing.
Mouse Met mRNA was detectable in liver and kidney of WT mice (EGFR+/+MET+/+) and B-hEGFR/hMET mice (EGFRH/HMETH/+). Human MET mRNA was detectable only in B-hEGFR/hMET mice (EGFRH/HMETH/+) but not in WT mice (EGFR+/+MET+/+). The positive band was confirmed to be correct by sequencing.Protein expression analysis:
MET and EGFR were detectable in liver and kidney of WT mice (EGFR+/+MET+/+) and B-hEGFR/hMET mice (EGFRH/HMETH/+) due to the cross-reactivity of antibodies.
