Basic Information
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Gene targeting strategy
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Gene targeting strategy for B-hFGL2 mice. The exons 1-2 of mouse Fgl2 gene that encode the full-length protein were replaced by human FGL2 exons 1-2 in B-hFGL2 mice.
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mRNA expression analysis
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Species-specific FGL2 gene expression analysis in wild-type and humanized B-hFGL2 mice by RT-PCR. Murine Fgl2 mRNA was detected in peritoneal exudate macrophages isolated from wild-type (+/+) mice, while human FGL2 mRNA was detected in homozygous B-hFGL2 (H/H) mice.
Strain specific FGL2 expression analysis in homozygous B-hFGL2 mice by western blot. Peritoneal exudative macrophages and splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hFGL2 mice (H/H) sitimulated by mIFNγ 20ng/ml in vitro for 24h, and analyzed by western blot with anti-FGL2 antibody. FGL2 was detectable in wild type mice and homozygous B-hFGL2 mice, as the antibody is crossly reactive with FGL2 in human and mice.
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Summary
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mRNA and Protein expression analysis:
Mouse Fgl2 mRNA was detectable in peritoneal exudative macrophages of wild-type (+/+). Human FGL2 mRNA was detectable in B-hFGL2 mice (H/H) but not in wild-type mice, and we proved that the protein could be expressed normally by western blot analysis.
