Basic Information
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Gene Targeting Strategy
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The human NT5E whole coding sequence was inserted following the 5’UTR of murine Nt5e in B-hCD73/hCD39 mice. A chimeric coding sequence of human ENTPD1 gene encoding the extracellular region and the murine Entpd1 gene encoding the second transmembrane and cytoplasmic region were inserted in exon 2 where it is after the first transmembrane region of mouse Entpd1 gene in B-hCD73/hCD39 mice. Meanwhile the sequences of mouse Entpd1 gene encoding the extracellular region on exon 2 and the following sequences on part of intron 2-3 were replaced with the chimeric coding sequences.
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Protein Expression Analysis
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Species-specific CD73 and CD39 expression analysis in double humanized B-hCD73/hCD39 mice. Splenocytes were collected from wild-type C57BL/6 (+/+) and homozygous B-hCD73/hCD39 (H/H) mice and analyzed by flow cytometry using species-specific anti-CD39 antibodies. Human CD73 and CD39 was exclusively detected in B-hCD73/hCD39 mice compared to wild-type mice.