Basic Information
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Targeting strategy
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Gene targeting strategy for B-hTNFR2/hTNFR1/hLTBR mice.
The exons 2~6 of mouse Tnfr2 gene that encode the extracellular domain were replaced by human TNFR2 exons 2~6 in B-hTNFR2/hTNFR1/hLTBR mice. The exons 2~6 of mouse Tnfr1 gene that encode the extracellular domain were replaced by human TNFR1 exons 2~6 in B-hTNFR2/hTNFR1/hLTBR mice. The exons 1-7 of mouse Ltbr gene that encode the extracellular domain were replaced by human LTBR exons 1-7 in B-hTNFR2/hTNFR1/hLTBR mice.
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TNFR1 protein expression analysis in granulocytes
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Species-specific TNFR1 protein expression analysis in wild-type and humanized B-hTNFR2/hTNFR1/hLTBR mice. Splenocytes were isolated from wild-type C57BL/6 (+/+) and homozygous B-hTNFR2/hTNFR1/hLTBR (H/H) mice and analyzed by flow cytometry using species-specific anti-TNFR1 antibodies. Mouse TNFR1 protein was detected in wild-type mice, while human TNFR1 protein was detected in B-hTNFR2/hTNFR1/hLTBR mice.
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LTBR protein expression analysis in macrophages
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Species-specific LTBR protein expression analysis in wild-type and humanized B-hTNFR2/hTNFR1/hLTBR mice. Spleen, blood and bone marrow cells were isolated from wild-type C57BL/6 (+/+) and homozygous B-hTNFR2/hTNFR1/hLTBR (H/H) mice, and analyzed by flow cytometry using species-specific anti-LTBR antibodies. Mouse LTBR protein was detected in wild-type mice, while human LTBR protein was detected in B-hTNFR2/hTNFR1/hLTBR mice.
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TNFR2 protein expression analysis in regulatory T cells (Tregs)
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Species-specific TNFR2 protein expression analysis in wild-type and humanized B-hTNFR2/hTNFR1/hLTBR mice. Following anti-CD3ε stimulation in vivo, splenocytes were isolated from wild-type C57BL/6 (+/+) and homozygous B-hTNFR2/hTNFR1/hLTBR (H/H) mice, and analyzed by flow cytometry using species-specific anti-TNFR2 antibodies. Mouse TNFR2 protein was detected in wild-type mice, while human TNFR2 protein was detected in B-hTNFR2/hTNFR1/hLTBR mice.
