Gene targeting strategy for B-hCTLA4/hLAG3 mice. The exon 2 of mouse Ctla4 gene that encodes the extracellular domain was replaced by human CTLA4 exon 2 in B-hCTLA4/hLAG3 mice. The exons 2-3 of mouse Lag3 gene that encode the extracellular domain were replaced by human LAG3 exons 2-3 in B-hCTLA4/hLAG3 mice.
Protein Expression Analysis
Strain specific CTLA4 and LAG3 expression analysis in homozygous B-hCTLA4/hLAG3 mice by flow cytometry. Splenocytes were collected from WT and homozygous B-hCTLA4/hLAG3 (H/H) mice stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with species-specific anti-CTLA4 and anti-LAG3 antibody. Mouse CTLA4 was detectable in WT mice, mouse LAG3 was detectable in homozygous B-hCTLA4/hLAG3 and WT mice due to the anti-mouse LAG3 antibody cross-reacts with human LAG3. Human CTLA4 and LAG3 were exclusively detectable in homozygous B-hCTLA4/hLAG3 but not WT mice.
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