Basic Information
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Targeting strategy
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Gene targeting strategy for B-hBTLA/hHVEM mice.
The exon 2 of mouse Btla gene that encodes the extracellular domain was replaced by human BTLA exon 2 in B-hBTLA/hHVEM mice. The exons 1~6 of mouse Tnfrsf14 gene that encode the extracellular domain were replaced by human TNFRSF14 exons 1~6 in B-hBTLA/hHVEM mice.
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mRNA expression analysis
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Strain specific analysis of BTLA and HVEM gene expression in wild type (WT) mice and B-hBTLA/hHVEM mice by RT-PCR.
Mouse Btla and Hvem mRNA was detectable only in splenocytes of WT mice (+/+). Human BTLA and HVEM mRNA was detectable only in homozygous B-hBTLA/hHVEM mice (H/H;H/H) but not in WT mice (+/+).
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Protein expression analysis in B cells
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Strain specific BTLA expression analysis in homozygous B-hBTLA/hHVEM mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hBTLA/hHVEM mice (H/H;H/H), and analyzed by flow cytometry with species-specific anti-BTLA antibody. Mouse BTLA was detectable in WT mice (+/+). Human BTLA was exclusively detectable in homozygous B-hBTLA/hHVEM mice (H/H;H/H) but not in WT mice (+/+).
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Protein expression analysis in T cells
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Strain specific HVEM expression analysis in homozygous B-hBTLA/hHVEM mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hBTLA/hHVEM mice (H/H;H/H), and analyzed by flow cytometry with species-specific anti-HVEM antibody. Mouse HVEM was detectable in WT mice (+/+). Human HVEM was exclusively detectable in homozygous B-hBTLA/hHVEM mice (H/H;H/H) but not in WT mice (+/+).
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Summary
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mRNA expression analysis:
Mouse Btla and Hvem mRNA was detectable only in splenocytes of WT mice (+/+). Human BTLA and HVEM mRNA was detectable only in homozygous B-hBTLA/hHVEM mice (H/H;H/H) but not in WT mice (+/+).
Protein expression analysis:
Human BTLA was exclusively detectable on B cells of homozygous B-hBTLA/hHVEM mice (H/H;H/H) but not in WT mice (+/+), and mouse BTLA was detectable only in WT mice (+/+).
Human HVEM was exclusively detectable on T cells of homozygous B-hBTLA/hHVEM mice (H/H;H/H) but not in WT mice (+/+), and mouse HVEM was detectable only in WT mice (+/+).
