Gene targeting strategy for B-hLY6E mice. The exons 3-4 of mouse Ly6e gene that encode the extracellular domain were replaced by human LY6E exons 3-4 in B-hLY6E mice.

Strain specific analysis of LY6E gene expression in wild-type mice and B-hLY6E mice by RT-PCR. Lung and spleen RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hLY6E mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human LY6E primers. Mouse Ly6e mRNA was only detectable in wild-type mice, human LY6E mRNA was exclusively detectable in homozygous B-hLY6E mice but not in wild-type mice.
Strain specific LY6E expression analysis in homozygous B-hLY6E mice by western blot. Kidney, liver, brain, spleen and lung tissue were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hLY6E mice(H/H), and analyzed by western blot with anti-LY6E antibody. Human LY6E was detectable in wild-type mice and homozygous B-hLY6E mice, as the antibody is crossly reactive with LY6E in human and mice.

Strain specific LY6E expression analysis in homozygous B-hLY6E mice by western blot. Kidney, liver, brain, spleen and lung tissue were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hLY6E mice (H/H), analyzed by western blot with anti-LY6E antibody (LSBio, LSB15888). LY6E was detectable in wild-type mice and homozygous B-hLY6E mice, as the antibody is crossly reactive with LY6E in human and mouse.

Strain specific LY6E expression analysis in wild-type C57BL/6JNifdc mice and homozygous B-hLY6E mice (H/H) by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hLY6E mice (H/H). Protein expression was analyzed with anti LY-6E antibody (BD, 560654) by flow cytometry. LY6E was detectable on B cells, T cells, CD4+ T cells and CD8+ T cells from splenocytes of homozygous B-hLY6E mice and wild-type C57BL/6JNifdc mice. The antibody was cross-reactive between human and mouse.

Strain specific LY6E expression analysis in wild-type C57BL/6JNifdc mice and homozygous B-hLY6E mice (H/H) by flow cytometry. Thymus cells were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hLY6E mice (H/H). Protein expression was analyzed with anti LY-6E antibody (BD, 560654) by flow cytometry. LY6E was detectable on CD4+ T cells, CD8+ T cells, CD4+/CD8+ T cells from thymus cells of homozygous B-hLY6E mice and wild-type C57BL/6JNifdc mice. The antibody was cross-reactive between human and mouse.