Basic Information
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Targeting strategy
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Gene targeting strategy for B-hSIGLEC8 mice. The BAC containing the whole sequence of human SIGLEC8 gene was inserted into Rosa26 locus in B-hSIGLEC8 mice.
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Protein expression analysis
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SIGLEC8 protein expression analysis in humanized B-hSIGLEC8 mice by flow cytometry. Blood and peritoneal lavage fluid were collected from wild-type C57BL/6 (+/+) and homozygous B-hSIGLEC8 (H/H) mice, and analyzed by flow cytometry using an anti-hSIGLEC8 antibody. Human SIGLEC8 protein was detected in eosinophils (88%, mSiglec-F+,mCD11c-) and mast cells (99.8%, mFcεRIα+,mCD117(c-kit)+,CD11b-) in B-hSIGLEC8 mice.
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Eosinophils and mast cells in B-hSIGLEC8 mice bind to anti-human SIGLEC8 antibody
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SIGLEC8 protein expression analysis in eosinophils and mast cells from humanized B-hSIGLEC8 mice. Eosinophils and peritoneal lavage-mast cells isolated from wild-type C57BL/6 (+/+) and B-hSIGLEC8 (H/H) mice were analyzed by flow cytometry to assess human SIGLEC8 protein expression using the Benchmark antibody Lirentelimab (in house). Eosinophils were gated from mSiglec-F+,mCD11c- population, while mast cells were gated from CD45+mFcεRIα+,mCD117(c-kit)+CD11b- population. Lirentelimab exclusively bound to human SIGLEC8 in B-hSIGLEC8 mice compared to wild-type mice.