Gene targeting strategy for B-hEVA-1 mice. The exons 2-4 of mouse Eva1 gene that encode the extracellular domain were replaced by human EVA-1 exons 2-4 in B-hEVA-1 mice.

Strain specific analysis of EVA-1 gene expression in wild-type mice and B-hEVA-1 mice by RT-PCR. Mouse Eva1 mRNA was detectable in thymus of wild-type mice (+/+). Human EVA-1 mRNA was detectable only in homozygous B-hEVA-1 mice(H/H) but not in wild-type mice. Strain specific EVA-1 expression analysis in homozygous B-hEVA-1 mice by flow cytometry. Granulocytes, macrophages, dendritic cells from blood were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hEVA-1 mice(H/H), and analyzed by flow cytometry with anti-EVA-1 antibody. Human EVA-1 was detectable in wild-type mice and homozygous B-hEVA-1 mice, as the antibody is crossly reactive with EVA-1 in human and mice.

Strain specific EVA-1 expression analysis in homozygous B-hEVA-1 mice by flow cytometry. DN3 cells, epithelial cells from thymus were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hEVA-1 mice(H/H), and analyzed by flow cytometry with anti-EVA-1 antibody. EVA-1 was detectable only in wild-type mice but not in B-hEVA-1 mice .