Basic Information
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Gene targeting strategy
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Gene targeting strategy for B-hPD-1/hPD-L1/hVEGFR2 mice. The exon 2 of mouse PD-1 gene that encodes the IgV domain was replaced by human PD-1 exon 2 in B-hPD-1/hPD-L1/hVEGFR2 mice. The exon 3 of mouse pdl1 gene that encodes the IgV domain was replaced by human PD-L1 exon 3 in B-hPD-1/hPD-L1/hVEGFR2 mice. The exons 2-15 of mouse Vegfr2 gene that encode the extracellular region were replaced by human VEGFR2 exons 2-15 in B-hPD-1/hPD-L1/hVEGFR2 mice.
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Protein expression analysis
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Strain specific PD-1 expression analysis in wild-type C57BL/6 and homozygous B-hPD-1/hPD-L1/hVEGFR2 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hPD-1/hPD-L1/hVEGFR2 mice (H/H) after stimulated with anti-CD3ε in vivo or not. Mouse PD-1 was detectable on CD4+ T cells and CD8+ T cell in wild-type C57BL/6 mice. Human PD-1 was exclusively detectable on CD4+ T cells and CD8+ T cell in homozygous B-hPD-1/hPD-L1/hVEGFR2 mice but not in wild-type mice.
Strain specific PD-L1 expression analysis in wild-type C57BL/6 and homozygous B-hPD-1/hPD-L1/hVEGFR2 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hPD-1/hPD-L1/hVEGFR2 mice (H/H) after stimulated with anti-CD3ε in vivo or not. Mouse PD-L1 was detectable on CD4+ T cells and CD8+ T cell in wild-type C57BL/6 mice. Human PD-L1 was exclusively detectable on CD4+ T cells and CD8+ T cell in homozygous B-hPD-1/hPD-L1/hVEGFR2 mice but not in wild-type mice.
Strain specific VEGFR2 expression analysis in wild-type C57BL/6 and homozygous B-hPD-1/hPD-L1/hVEGFR2 mice by flow cytometry. Lung endothelial cells were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hPD-1/hPD-L1/hVEGFR2 mice (H/H). Mouse VEGFR2 was detectable in lung endothelial cells in wild-type C57BL/6 mice. Human VEGFR2 was only detectable in homozygous B-hPD-1/hPD-L1/hVEGFR2 mice but not in wild-type mice.