Basic Information
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Targeting strategy
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Gene targeting strategy for B-hFGL1/hLAG3 plus mice.
The exons 2-8 of mouse Fgl1 gene that encode the full-length protein were replaced by human FGL1 exons 2~8 in B-hFGL1/hLAG3 plus mice.
The exons 2-7 of mouse Lag3 gene that encode the extracellular domain were replaced by human LAG3 exons 2-7 in B-hFGL1/hLAG3 plus mice.
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mRNA expression analysis
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Strain specific analysis of FGL1 gene expression in wild-type C57BL/6 mice and B-hFGL1/hLAG3 plus mice by RT-PCR. Mouse Fgl1 mRNA was detectable only in liver of wild-type C57BL/6 mice (+/+). Human FGL1 mRNA was detectable only in homozygous B-hFGL1/hLAG3 plus mice (H/H; H/H), but not in wild-type mice.
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Protein expression analysis
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Strain specific FGL1 expression analysis in homozygous B-hFGL1/hLAG3 plus mice by ELISA. Plasma was collected from wild-type C57BL/6 mice (+/+) and homozygous B-hFGL1/hLAG3 plus mice (H/H;H/H), and analyzed by ELISA with species-specific FGL1 ELISA kit. Human FGL1 was exclusively detectable in homozygous B-hFGL1/hLAG3 plus mice (H/H;H/H) but not wild-type mice.
Strain specific LAG3 expression analysis in homozygous B-hFGL1/hLAG3 plus mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 (+/+) and homozygous B-hFGL1/hLAG3 plus mice (H/H; H/H) stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with species-specific anti-LAG3 antibody. Mouse LAG3 was detectable in C57BL/6 (+/+) . Human LAG3 was exclusively detectable in homozygous B-hFGL1/hLAG3 plus mice (H/H; H/H) but not WT mice.
