Basic Information

Common name
B-hMASP2 mice
Background
C57BL/6N
Catalog number
110872
Aliases
MASP2,Map19
NCBI Gene ID

Targeting strategy

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.

Protein expression analysis

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.

MASP2 expression pattern analysis by IHC

Representative human MASP2 expression in different tissues of B-hMASP2 mice by IHC. Tissues were stained with antibodies for MASP2 (A-F) or anti-IgG antibody (G). Liver and kidney show MASP2 positive both in B-hMASP2 mice (C-D) and C57BL/6 mice (A-B). Human CDX HepG2 as a positive control (E); Dog tissue was a negative control (F); Mouse liver tissues from the wild-type C57BL/6 mice (G). Original magnification ×200. Abbreviations: IHC, immunohistochemistry.

mRNA expression analysis

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.

Analysis of leukocyte cell subpopulations in spleen

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.

Analysis of leukocyte cell subpopulations in lymph node

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.

Analysis of leukocyte cell subpopulations in blood

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.

Summary

Protein expression analysis:

Human IL18BP was exclusively detectable in homozygous B-hIL18BP mice but not in wild-type mice.

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