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).
