Basic Information
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Targeting strategy
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Gene targeting strategy for B-hCD73/h4-1BB mice. The exons 2-7 of mouse Tnfrsf gene that encode the extracellular domain were replaced by human TNFRSF exons 2-7 in B-hCD73/h4-1BB mice. The full encoding region sequences of human NT5E gene was insert into mouse Nt5e in B-hCD73/h4-1BB mice.
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Protein expression analysis
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Strain specific CD73 expression analysis in wild-type C57BL/6 mice and homozygous B-hCD73/h4-1BB mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice and homozygous B-hCD73/h4-1BB mice, stimulated with anti-CD3ε in vivo (7.5 μg/mice for 24 hours, i.p.). Mouse CD73 was only detectable in wild-type mice. Human CD73 was only detectable in homozygous B-hCD73/h4-1BB mice.
Strain specific CD73 expression analysis in wild-type C57BL/6 mice and homozygous B-hCD73/h4-1BB mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice and homozygous B-hCD73/h4-1BB mice, stimulated with anti-CD3ε in vivo (7.5 μg/mice for 24 hours, i.p.). Mouse CD73 was only detectable in wild-type mice. Human CD73 was only detectable in homozygous B-hCD73/h4-1BB mice.
Strain specific CD73 expression analysis in wild-type C57BL/6 mice and homozygous B-hCD73/h4-1BB mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice and homozygous B-hCD73/h4-1BB mice, stimulated with anti-CD3ε in vivo (7.5 μg/mice for 24 hours, i.p.). Mouse CD73 was only detectable in wild-type mice. Human CD73 was only detectable in homozygous B-hCD73/h4-1BB mice.
Strain specific 4-1BB expression analysis in wild-type C57BL/6 mice and homozygous B-hCD73/h4-1BB mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice and homozygous B-hCD73/h4-1BB mice, stimulated with anti-CD3ε in vivo (7.5 μg/mice for 24 hours, i.p.). Mouse 4-1BB was only detectable in wild-type mice. Human 4-1BB was only detectable in homozygous B-hCD73/h4-1BB mice.
Strain specific 4-1BB expression analysis in wild-type C57BL/6 mice and homozygous B-hCD73/h4-1BB mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice and homozygous B-hCD73/h4-1BB mice, stimulated with anti-CD3ε in vivo (7.5 μg/mice for 24 hours, i.p.). Mouse 4-1BB was only detectable in wild-type mice. Human 4-1BB was only detectable in homozygous B-hCD73/h4-1BB mice.
Strain specific 4-1BB expression analysis in wild-type C57BL/6 mice and homozygous B-hCD73/h4-1BB mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice and homozygous B-hCD73/h4-1BB mice, stimulated with anti-CD3ε in vivo (7.5 μg/mice for 24 hours, i.p.). Mouse 4-1BB was only detectable in wild-type mice. Human 4-1BB was only detectable in homozygous B-hCD73/h4-1BB mice.
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Summary
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Protein expression analysis:
Human CD73 and 4-1BB were exclusively detectable in homozygous B-hCD73/h4-1BB mice but not in wild type mice.