Gene targeting strategy for B-hCLEC9A mice. The exons 2~6 of mouse Clec9a gene that encode the extracellular region were replaced by human CLEC9A exons 5~9 in B-hCLEC9A mice.  

Strain specific CLEC9A expression analysis in homozygous B-hCELC9A mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCLEC9A mice (H/H), and analyzed by flow cytometry with species-specific anti-CLEC9A antibody. Mouse CLEC9A was detectable in mCD8α+ DCs and mCD103+ DCs of wild-type mice. Human CLEC9A was exclusively detectable in mCD8α+ DCs and mCD103+ DCs of homozygous B-hCLEC9A mice but not in wild-type mice.

Frequency of leukocyte subpopulations in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 mice (female, n=3, 6-week-old) and homozygous B-hCLEC9A mice (female, n=3, 6-week-old). A. Flow cytometry analysis of the splenocytes was performed to assess the frequency of leukocyte subpopulations. B. Frequency of T cell subpopulations. Percentages of T cells, B cells, NK cells, dendritic cells, neutrophils, monocytes, macrophages, CD4+ T cells, CD8+ T cells and Tregs in B-hCLEC9A mice were similar to those in C57BL/6N mice. Values are expressed as mean ± SEM.