Basic Information
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Targeting strategy
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Gene targeting strategy for B-hMASP2 mice. The full coding sequence of human MASP2 was inserted into mouse Masp2 gene locus in B-hMASP2 mice, so the mouse Masp2 gene was disrupted.
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Protein expression analysis
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Strain specific MASP2 expression analysis in heterozygous B-hMASP2 mice by ELISA. Serum were collected from C57BL/6 and heterozygous B-hMASP2 mice, and analyzed by ELISA with species-specific MASP2 ELISA kit. Mouse MASP2 was detectable in C57BL/6 and B-hMASP2 mice. Human MASP2 was exclusively detectable in B-hMASP2 mice but not in C57BL/6 mice.
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mRNA expression analysis
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Strain specific analysis of MASP2 gene expression in wild-type C57BL/6 mice and hMASP2 mice by RT-PCR. Human MASP2 mRNA was detectable in homozygous B-hMASP2 mice (H/H), but not in wild-type C57BL/6 mice (+/+). Mouse MASP2 mRNA was both detectable in wild-type C57BL/6 and homozygous B-hMASP2 mice.
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Analysis of leukocyte cell subpopulations in spleen
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Analysis of spleen leukocyte subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hMASP2 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 T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hMASP2 mice were similar to those in the C57BL/6 mice, demonstrating that MASP2 humanized does not change the overall development, differentiation or distribution of these cell types in spleen.
Analysis of spleen T cell subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hMASP2 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 TCRβ+ 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-hMASP2 mice were similar to those in the C57BL/6 mice, demonstrating that MASP2 humanized does not change the overall development, differentiation or distribution of these cell types in spleen.
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Analysis of leukocyte cell subpopulations in lymph node
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Analysis of lymph node leukocyte subpopulations by FACS. Lymph nodes were isolated from female C57BL/6 and B-hMASP2 mice (n=3, 6-week-old). Flow cytometry analysis of the lymph node 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 T cells, B cells, NK cells in homozygous B-hMASP2 mice were similar to those in the C57BL/6 mice, demonstrating that MASP2 humanized does not change the overall development, differentiation or distribution of these cell types in lymph node.
Analysis of lymph node T cell subpopulations by FACS. Lymph nodes were isolated from female C57BL/6 and B-hMASP2 mice (n=3, 6-week-old). Flow cytometry analysis of the lymph node was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for TCRβ+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-hMASP2 mice were similar to those in the C57BL/6 mice, demonstrating that MASP2 humanized does not change the overall development, differentiation or distribution of these cell types in lymph node.
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Analysis of leukocyte cell subpopulations in blood
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Analysis of blood leukocyte subpopulations by FACS. Blood cells were isolated from female C57BL/6 and B-hMASP2 mice (n=3, 6-week-old). Flow cytometry analysis of the blood 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 T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hMASP2 mice were similar to those in the C57BL/6 mice, demonstrating that MASP2 humanized does not change the overall development, differentiation or distribution of these cell types in blood.
Analysis of blood T cell subpopulations by FACS. Blood cells were isolated from female C57BL/6 and B-hMASP2 mice (n=3, 6-week-old). Flow cytometry analysis of the blood was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for TCRβ+ 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-hMASP2 mice were similar to those in the C57BL/6 mice, demonstrating that MASP2 humanized does not change the overall development, differentiation or distribution of these cell types in blood.
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Summary
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Protein expression analysis:
Human IL18BP was exclusively detectable in homozygous B-hIL18BP mice but not in wild-type mice.
