Basic Information
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Details
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1.Generation of gene editing mouse model and expression analysis
RT-PCR analysis
RT-PCR analysis of B-hIL10 mice
The hIL10 mRNA, but not mIl10 mRNA was detectable in splenocytes of the homozygous B-hIL10 mice .
Protein expression analysis
Serum from both wild type (WT) C57BL/6 and homozygous B-hIL10 mice were analyzed by ELISA.
When treated with LPS, mouse IL10 was only detectable in WT C57BL/6 mice, while human IL10 was only detectable in homozygous B-hIL10 mice. n=3.
2.Functional analysis
Analysis of leukocytes cell subpopulation in B-hIL10 mice
Analysis of leukocytes subpopulation in spleen
Splenocytes were isolated from C57BL/6 and B-hIL10 mice (n=3). The proportion of leukocytes subpopulation was tested by flow cytometry. As a result, the expression profile of leukocytes subpopulation in homozygous B-hIL10 mice is similar to that in the C57BL/6 mice, indicating that differentiation of T, B, NK, Monocyte, DC and macrophage cells are not affected by the humanization of hIL10.
Analysis of T cell subpopulation in B-hIL10 mice
Analysis of T cell subpopulation in spleen
The lymphocytes were isolated from spleen in C57BL/6 and B-hIL10 mice (n=3). The proportion of T cells subpopulation was tested by flow cytometry. As a result, the expression profile of lymphocyte subpopulation in homozygous B-hIL10 mice is similar to that in the C57BL/6 mice, indicating that T cell differentiation is not affected by the humanization of hIL10.
Analysis of leukocytes cell subpopulation in B-hIL10 mice
Analysis of leukocytes subpopulation in spleen
Blood were isolated from C57BL/6 and B-hIL10 mice (n=3). The proportion of leukocytes subpopulation was tested by flow cytometry. As a result, the expression profile of leukocytes subpopulation in homozygous B-hIL10 mice is similar to that in the C57BL/6 mice, indicating that differentiation of T, B, NK, Monocyte, DC and macrophage cells are not affected by the humanization of hIL10.
Analysis of T cell subpopulation in B-hIL10 mice
Analysis of T cell subpopulation in spleen
The lymphocytes were isolated from Blood in C57BL/6 and B-hIL10 mice (n=3). The proportion of T cells subpopulation was tested by flow cytometry. As a result, the expression profile of lymphocyte subpopulation in homozygous B-hIL10 mice is similar to that in the C57BL/6 mice, indicating that T cell differentiation is not affected by the humanization of hIL10.
