Gene targeting strategy for B-hSIGLEC10 mice. The exons 3~10 of mouse Siglecg gene that encode the extracellular domain were replaced by human SIGLEC10 exons 3~10 in B-hSIGLEC10 mice.

Strain specific SIGLEC10 expression analysis in homozygous B-hSIGLEC10 mice by flow cytometry. Splenocytes were collected from WT and homozygous B-hSIGLEC10 (H/H) mice, and analyzed by flow cytometry with species-specific SIGLEC10 antibody. Mouse SIGLEC10 was detectable in WT mice. Human SIGLEC10 was exclusively detectable in homozygous B-hSIGLEC10 mice but not in WT mice.

Strain specific SIGLEC10 expression analysis in homozygous B-hSIGLEC10 mice by flow cytometry. Splenocytes were collected from WT and homozygous B-hSIGLEC10 (H/H) mice, and analyzed by flow cytometry with species-specific SIGLEC10 antibody. Mouse SIGLEC10 was detectable in WT mice. Human SIGLEC10 was exclusively detectable in homozygous B-hSIGLEC10 but not WT mice.

Analysis of spleen leukocyte subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hSIGLEC10 mice(n=3, 6-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hSIGLEC10 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hSIGLEC10 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in spleen. Values are expressed as mean ± SEM.

Analysis of spleen T cell subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hSIGLEC10 mice(n=3, 6-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD8+ T cells, CD4+ T cells, and Tregs in homozygous B-hSIGLEC10 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hSIGLEC10 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in spleen. Values are expressed as mean ± SEM.

Analysis of spleen leukocyte subpopulations by FACS. Lymph node were isolated from female C57BL/6 and B-hSIGLEC10 mice(n=3, 6-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hSIGLEC10 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hSIGLEC10 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in lymph node. Values are expressed as mean ± SEM.

Analysis of blood leukocyte subpopulations by FACS. Leukocytes were isolated from female C57BL/6 and B-hSIGLEC10 mice(n=3, 6-week-old) Flow cytometry analysis of the leukocytes was performed to assess leukocyte subpopulations. Representative FACS plots. Single live CD45+ cells were gated for TCRb+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD4+, CD8+, Tregs in homozygous B-Hsiglec10 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hSIGLEC10 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in blood. Values are expressed as mean ± SEM.