Gene targeting strategy for B-hCD63 mice.
The exons 2-8 of mouse Cd63 gene that encode the full-length protein were replaced by human CD63 exons 2-8 in B-hCD63 mice.

Strain specific analysis of CD63 gene expression in wild type and B-hCD63 mice by RT-PCR. Kidney RNA were isolated from wildtype C57BL/6 mice (+/+) and homozygous B-hCD63 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human CD63 primers. Mouse Cd63 mRNA was detectable only in wild type mice. Human CD63 mRNA was detectable only in B-hCD63 mice but not in wild type mice.

CD63 expression analysis in wild-type C57BL/6 mice and homozygous humanized B-hCD63 mice by flow cytometry. Bone marrow cells were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCD63 mice (H/H) (Female, 6-week-old, n=1). Protein expression was analyzed with anti-human CD63 antibody (Biolegend, 353007) and anti-mouse CD63 antibody (Biolegend, 143903) by flow cytometry. Human CD63 was exclusively detectable on monocytes and macrophages in the bone marrow of homozygous B-hCD63 mice, but not in wild-type C57BL/6 mice.