Basic Information

Strain name
C57BL/6-Tnfrsf9tm1(TNFRSF9)Erbb2tm1(ERBB2)/Bcgen
Common Name
B-h4-1BB/hHER2 mice
Background
C57BL/6
Catalog Number
111869
Aliases
TNFRSF9, TNF receptor superfamily member 9,4-1BB, CD137, CDw137, ILA; ERBB2, CD340, HER-2, HER-2/neu, HER2, MLN 19, NEU, NGL, TKR1, VSCN2
NCBI Gene ID

Targeting strategy

Gene targeting strategy for B-h4-1BB/hHER2 mice.

The exons 2-7 of mouse Tnfrsf9 gene that encode the extracellular domain were replaced by human TNFRSF9 exons 3-8 in B-h4-1BB/hHER2 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-h4-1BB/hHER2 mice.

mRNA expression analysis

Strain specific analysis of 4-1BB gene expression in wild type (WT) mice and B-h4-1BB/hHER2 mice  by RT-PCR.

Mouse Tnfrsf9 mRNA was detectable only in spleen of WT mice (+/+). Human TNFRSF9 mRNA was detectable only in homozygous B-h4-1BB/hHER2 mice  (H/H;H/H) but not in WT mice (+/+).

Strain specific analysis of HER2 gene expression in wild type (WT) mice and B-h4-1BB/hHER2 mice  by RT-PCR.

Mouse Erbb2 mRNA was detectable only in liver of WT mice (+/+). Human ERBB2 mRNA was detectable only in homozygous B-h4-1BB/hHER2 mice  (H/H;H/H) but not in WT mice (+/+).

Protein expression analysis

Strain specific 4-1BB expression analysis in homozygous B-h4-1BB/hHER2 mice by flow cytometry.

Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-h4-1BB/hHER2 mice (H/H;H/H) stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with species-specific anti-4-1BB antibody. Mouse 4-1BB was detectable in WT mice (+/+). Human 4-1BB was exclusively detectable in homozygous B-h4-1BB/hHER2 mice (H/H;H/H) but not in WT mice (+/+).

IHC analysis of HER2 expression

Immunohistochemical (IHC) analysis of HER2 expression in homozygous B-h4-1BB/hHER2 mice.

The liver and mammary gland were collected from WT mice and homozygous B-h4-1BB/hHER2 mice and analyzed by IHC with anti-HER2 antibody. HER2 was detectable in WT mice and homozygous B-h4-1BB/hHER2 mice due to the cross-reactivity of the antibody. The arrow indicates tissue cells with positive HER2 staining (brown).

Summary

mRNA expression analysis:

Mouse Tnfrsf9 mRNA was detectable only in spleen of WT mice (+/+). Human TNFRSF9 mRNA was detectable only in homozygous B-h4-1BB/hHER2 mice  (H/H;H/H) but not in WT mice (+/+).

Mouse Erbb2 mRNA was detectable only in liver of WT mice (+/+). Human ERBB2 mRNA was detectable only in homozygous B-h4-1BB/hHER2 mice  (H/H;H/H) but not in WT mice (+/+).

Protein expression analysis:

Human 4-1BB was exclusively detectable on T cells of homozygous B-h4-1BB/hHER2 mice  (H/H;H/H) but not in WT mice (+/+), and mouse 4-1BB/hHER2 was detectable only in WT mice (+/+).

HER2 was detectable in liver and mammary gland of WT mice (+/+) and homozygous B-h4-1BB/hHER2 mice (H/H;H/H) due to the cross-reactivity of antibodies.

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