Basic Information
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Gene Targeting Strategy
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Exon 2 of the murine Mertk gene was replaced by the human MERTK chimeric CDS in B-hMERTK mice.
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mRNA expression analysis
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MERTK gene expression analysis in wild-type and humanized B-hMERTK mice. MERTK mRNA expression level in homozygous B-hMERTK (H/H) mice was similar to that in wild-type (+/+) mice, demonstrating that introduction of hMERTK in place of its mouse counterpart does not change the overall expression level of MERTK mRNA.
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Protein expression analysis
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Species–specific MERTK protein expression analysis in humanized B-hMERTK mice. Peritoneal washes were collected from wild-type C57BL/6 (+/+) and heterozygous B-hMERTK (H/+) mice, and analyzed by flow cytometry using species-specific anti-MERTK antibodies. Human MERTK protein was exclusively detected in B-hMERTK mice compared to wild-type mice. -
Analysis of spleen immune cells
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Analysis of spleen leukocyte subpopulations by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 (+/+) and homozygous B-hMERTK (H/H) mice (male, n=3, 7-week-old), and analyzed by flow cytometry to assess leukocyte subpopulations. (A) Representative flow cytometry plots. Single live cells were gated on CD45+ cells and used for further analysis as indicated. (B) Percent of T cells, B cells, NK cells, CD4+ T cells, CD8+ T cells, dendritic cells, granulocytes, monocytes and macrophages in humanized B-hMERTK mice were similar to those in wild-type mice, demonstrating that hMERTK in place of its mouse counterpart does not change the overall development, differentiation or distribution of these spleen cell subtypes. Values are expressed as mean ± SEM.
Analysis of spleen T cell subpopulations by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 (+/+) and homozygous B-hMERTK (H/H) mice (male, n=3, 7-week-old), and analyzed by flow cytometry to assess leukocyte subpopulations. (A) Representative flow cytometry plots. Single live CD45+ cells were gated on T cells and used for further analysis as indicated. (B) Percent of CD8+ T cells, CD4+ T cells, and Tregs in humanized B-hMERTK mice were similar to those in wild-type mice, demonstrating that introduction of hMERTK in place of its mouse counterpart does not change the overall development, differentiation or distribution of these spleen T cell subtypes. Values are expressed as mean ± SEM.
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Analysis of lymph node immune cells
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Analysis of lymph node leukocyte subpopulations by flow cytometry. Leukocytes were isolated from wild-type C57BL/6 (+/+) and homozygous B-hMERTK (H/H) mice (male, n=3, 7-week-old), and analyzed by flow cytometry to assess leukocyte subpopulations. (A) Representative flow cytometry plots. Single live cells were gated on CD45+ and used for further analysis as indicated. (B) Percent of T cells, B cells, NK cells, CD4+ T cells and CD8+ T cells in humanized B-hMERTK mice were similar to those in wild-type mice, demonstrating that introduction of hMERTK in place of its mouse counterpart does not change the overall development, differentiation or distribution of these lymph node cell subtypes. Values are expressed as mean ± SEM.
Analysis of lymph node T cell subpopulations by flow cytometry. Leukocytes were isolated from wild-type C57BL/6 (+/+) and homozygous B-hMERTK (H/H) mice (male, n=3, 7- week-old), and analyzed by flow cytometry to assess leukocyte subpopulations. Representative flow cytometry plots. Single live CD45+ cells were gated on TCRb+ T cells and used for further analysis as indicated. (B) Percent of CD8+ T cells, CD4+ T cells, and Tregs in humanized B-hMERTK mice were similar to those in wild-type mice, demonstrating that introduction of hMERTK in place of its mouse counterpart does not change the overall development, differentiation or distribution of these lymph node cell subtypes. Values are expressed as mean ± SEM.
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Analysis of blood immune cells
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Analysis of blood leukocyte subpopulations by flow cytometry. Leukocytes were isolated from wild-type C57BL/6 and homozygous B-hMERTK (H/H) mice (male, n=3, 7-week-old), and analyzed by flow cytometry to assess leukocyte subpopulations. (A) Representative flow cytometry plots. Single live cells were gated on CD45+ and used for further analysis as indicated. (B) Percent of T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in humanized B-hMERTK mice were similar to those in wild-type mice, demonstrating that introduction of hMERTK in place of its mouse counterpart does not change the overall development, differentiation or distribution of these blood cell subtypes. Values are expressed as mean ± SEM.
Analysis of blood T cell subpopulations by flow cytometry. Blood was isolated from wild-type C57BL/6 (+/+) and homozygous B-hMERTK (H/H) mice (male, n=3, 7-week-old), and analyzed by flow cytometry to assess leukocyte subpopulations. (A) Representative flow cytometry plots. Single live CD45+ cells were gated on TCRb+ T cells and used for further analysis as indicated here. (B) Percent of CD4+, CD8+, and Tregs in humanized B-hMERTK mice were similar to those in wild-type mice, demonstrating that introduction of hMERTK in place of its mouse counterpart does not change the overall development, differentiation or distribution of these blood cell subtypes. Values are expressed as mean ± SEM.
