Basic Information
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Gene Targeting Strategy
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Gene targeting strategy for B-hTSLP/hTSLPR mice. The exons 1-5 of mouse Tslp gene that encode the full-length protein were replaced by human TSLP exons 1-4 in B-hTSLP/hTSLPR mice. The extracellular and transmembrane region of human TSLPR gene and the cytoplasmic region of mouse Tslpr gene were constructed into a chimeric CDS vector and inserted into the mouse exon 2. The targeted mice will express the chimeric TSLPR protein, while mouse TSLPR will no longer express.
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TSLP Protein Expression Analysis
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Species-specific TSLP expression analysis in heterozygous B-hTSLP mice by ELISA. Calcipotriol (MC903) was dissolved in ethanol and topically applied on ears of either wild-type C57BL/6 (+/+) or heterozygous B-hTSLP mice (H/+) for 3 days. Ear grinding supernatant was analyzed by ELISA. Murine TSLP protein was detected in wild-type and heterozygous B-hTSLP mice, while human TSLP was exclusively detected in heterozygous B-hTSLP mice.
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TSLPR mRNA Expression Analysis
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Species-specific TSLPR gene expression analysis in wild type and heterozygous B-hTSLP/hTSLPR mice. Murine Tslpr mRNA was detected in splenocytes isolated from wild-type C57BL/6 (+/+) and heterozygous B-hTSLP/hTSLPR (H/+;H/+) mice. Human TSLPR mRNA was exclusively detected in humanized B-hTSLP/hTSLPR mice.
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TSLPR Protein Expression Analysis
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Species-specific TSLPR expression analysis in heterozygous B-hTSLP/hTSLPR mice. Macrophages were collected from wild-type C57BL/6 (+/+) and heterozygous B-hTSLP/hTSLPR (H/+;H/+) mice, and analyzed by flow cytometry using species-specific anti-TSLPR antibodies. Murine TSLPR protein was detected in wild-type and heterozygous B-hTSLP/hTSLPR mice, while human TSLPR protein was exclusively detected in humanized B-hTSLP/hTSLPR mice.
Species-specific TSLPR expression analysis in heterozygous B-hTSLP/hTSLPR mice. Following LPS stimulation, dendritic cells isolated from the bone marrow of wild-type C57BL/6 (+/+) and heterozygous B-hTSLP/hTSLPR (H/+) mice were analyzed by flow cytometry using species-specific anti-TSLPR antibodies. Murine TSLPR protein was detected in wild-type and heterozygous B-hTSLP/hTSLPR mice, while human TSLPR protein was exclusively detected in humanized B-hTSLP/hTSLPR mice.
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Poster
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Functional Analysis of Anti-TSLP Antibodies Using Humanized B-hTSLP/hTSLPR Mice