Basic Information
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Targeting strategy
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Gene targeting strategy for B-hHER2/hTROP2 mice. The exons 2-17 of mouse Erbb2 gene that encode the extracellular and transmembrane domains were replaced by human ERBB2 exons 2-17 in B-hHER2/hTROP2 mice. The exon 1 of mouse Trop2 gene that encodes the full-length protein was replaced by human TROP2 exon 1 in B-hHER2/hTROP2 mice.
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mRNA expression analysis
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Species-specific HER2 and TROP2 gene expression analyses in wild-type and humanized B-hHER2/hTROP2 mice by RT-PCR. Mouse Her2 and Trop2 mRNAs were detected in kidney tissue isolated from wild-type C57BL/6 (+/+) mice, while human HER2 and TROP2 mRNAs were detected in homozygous B-hHER2/hTROP2 (H/H) mice.
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Protein expression analysis
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Species-specific TROP2 protein expression analysis in wild-type and humanized B-hHER2/hTROP2 mice by western blot. Kidney tissue was isolated from wild-type C57BL/6 (+/+) and homozygous B-hHER2/hTROP2 (H/H) mice, and analyzed by western blot using an anti-TROP2 antibody. TROP2 protein was detected in wild-type and B-hHER2/hTROP2 mice due to the cross-reactive antibody.
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IHC analysis of HER2 expression
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Immunohistochemical (IHC) analysis of HER2 expression in wild-type and humanized B-hHER2/hTROP2 mice. Liver tissue was isolated from wild-type C57BL/6 (+/+) and homozygous B-hHER2/hTROP2 (H/H) mice and analyzed by IHC using an anti-HER2 antibody. HER2 protein was detected in wild-type and B-hHER2/hTROP2 mice due to the cross-reactive antibody. The arrow indicates tissue cells with positive HER2 staining (brown).
