Gene targeting strategy for B-hCD164 mice.
The exons 1-6 of mouse Cd164 gene that encode signal peptide, extracellular domain, transmembrane domain, cytoplasmic region and 3’UTR are replaced by human counterparts in B-hCD164 mice. The promoter and 5’UTR region of the mouse gene are retained. The human CD164 expression is driven by endogenous mouse Cd164 promoter, while mouse Cd164 gene transcription and translation will be disrupted.

Strain specific CD164 expression analysis in wild-type C57BL/6JNifdc mice and homozygous humanized B-hCD164 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hCD164 mice (H/H). Protein expression was analyzed with anti-human CD164 antibody (Biolegend, 324808) by flow cytometry. Human CD164 was exclusively detectable in homozygous B-hCD164 mice, but not in wild-type C57BL/6 mice.

Strain specific CD164 expression analysis in wild-type C57BL/6JNifdc mice and homozygous humanized B-hCD164 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hCD164 mice (H/H). Protein expression was analyzed with anti-human CD164 antibody (Biolegend, 324808) by flow cytometry. Human CD164 was exclusively detectable in homozygous B-hCD164 mice, but not in wild-type C57BL/6 mice.

Strain specific CD164 expression analysis in wild-type C57BL/6JNifdc mice and homozygous humanized B-hCD164 mice by flow cytometry. BM were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hCD164 mice (H/H). Protein expression was analyzed with anti-human CD164 antibody (Biolegend, 324808) by flow cytometry. Human CD164 was exclusively detectable in homozygous B-hCD164 mice, but not in wild-type C57BL/6 mice.

Strain specific CD164 expression analysis in wild-type C57BL/6JNifdc mice and homozygous humanized B-hCD164 mice by flow cytometry. BM were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hCD164 mice (H/H). Protein expression was analyzed with anti-human CD164 antibody (Biolegend, 324808) by flow cytometry. Human CD164 was exclusively detectable in homozygous B-hCD164 mice, but not in wild-type C57BL/6 mice.

Strain specific CD164 expression analysis in wild-type C57BL/6JNifdc mice and homozygous humanized B-hCD164 mice by flow cytometry. Blood cells were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hCD164 mice (H/H). Protein expression was analyzed with anti-human CD164 antibody (Biolegend, 324808) by flow cytometry. Human CD164 was exclusively detectable in homozygous B-hCD164 mice, but not in wild-type C57BL/6 mice.

Strain specific CD164 expression analysis in wild-type C57BL/6JNifdc mice and homozygous humanized B-hCD164 mice by flow cytometry. Blood cells were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hCD164 mice (H/H). Protein expression was analyzed with anti-human CD164 antibody (Biolegend, 324808) by flow cytometry. Human CD164 was exclusively detectable in homozygous B-hCD164 mice, but not in wild-type C57BL/6 mice.

Strain specific analysis of CD164 mRNA expression in wild-type C57BL/6JNifdc mice and B-hCD164 mice by RT-PCR. Spleen RNA was isolated from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hCD164 mice (H/H), and then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse and human Cd164 primers. Mouse Cd164 mRNA was only detectable in wild-type mice but not in homozygous B-hCD164 mice. Human CD164 mRNA was only detectable in homozygous B-hCD164 mice but not in wild-type mice.