Basic Information
-
Gene targeting strategy
-
Gene targeting strategy for B-hSIGLEC10 mice. Exons 3~10 of mouse Siglecg gene that encode the extracellular domain were replaced by human SIGLEC10 exons 3~10 in B-hSIGLEC10 mice.
-
Protein expression analysis
-
Protein expression analysis in B cells
Species-specific SIGLEC10 protein expression analysis in humanized B-hSIGLEC10 mice. Splenocytes were isolated from wild-type C57BL/6 (+/+) and heterozygous B-hSIGLEC10 (H/+) mice, and analyzed by flow cytometry using species-specific SIGLEC10 antibodies. Murine SIGLEC10 protein was detected in wild-type and heterozygous B-hSIGLEC10 mice, while human SIGLEC10 protein was exclusively detected in humanized B-hSIGLEC10 mice.
Protein expression analysis in macrophages
Species-specific SIGLEC10 protein expression analysis in humanized B-hSIGLEC10 mice. Splenocytes were isolated from wild-type C57BL/6 (+/+) and heterozygous B-hSIGLEC10 (H/+) mice, and analyzed by flow cytometry using species-specific SIGLEC10 antibodies. Murine SIGLEC10 protein was detected in wild-type and heterozygous B-hSIGLEC10 mice, while human SIGLEC10 protein was exclusively detected in humanized B-hSIGLEC10 mice.
-
Analysis of spleen leukocyte subpopulations
-
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
-
Analysis of spleen T cell subpopulations. 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.
