Basic Information
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Gene targeting strategy
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Gene targeting strategy for B-hTROP2 mice. Exon 1 of the mouse Trop2 gene that encodes the full-length protein was replaced by human TROP2 exons 1 in B-hTROP2 mice.
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mRNA expression analysis
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Species-specific TROP2 gene expression analysis in wild-type and humanized B-hTROP2 mice by RT-PCR. Mouse Trop2 mRNA was detected in skin tissue isolated from wild-type C57BL/6 (+/+) mice, while human TROP2 mRNA was detected in homozygous B-hTROP2 (H/H) mice.
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Protein expression analysis
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Species-specific TROP2 protein expression analysis in wild-type and humanized B-hTROP2 mice. Skin and kidney tissues were isolated from wild-type C57BL/6 (+/+) and homozygous B-hTROP2 (H/H) mice, and analyzed by western blot using anti-TROP2 antibodies. Mouse/human TROP2 protein was detected in wild-type mice due to the cross-reactive antibody, while human TROP2 protein was detected in B-hTROP2 mice and B-hTROP2 MC38 cells.
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Analysis of spleen immune cells in B-hTROP2 mice
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Analysis of spleen leukocyte subpopulations. Splenocytes were isolated from wild-type C57BL/6 and B-hTROP2 mice (female, n=3, 6 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, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hTROP2 mice were similar to those in wild-type mice, demonstrating that introduction of hTROP2 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these spleen cell types. Values are expressed as mean ± SEM.
Analysis of spleen T cell subpopulations. Splenocytes were isolated from wild-type C57BL/6 and B-hTROP2 mice (female, n=3, 6 week-old), and analyzed by flow cytometry to assess leukocyte subpopulations. (A) Representative flow cytometry plots. Single live CD45+ cells were gated on CD3 T cells and used for further analysis as indicated. (B) Percent of CD8+ T cells, CD4+ T cells and Treg cells in homozygous B-hTROP2 mice were similar to those in wild-type mice, demonstrating that introduction of hTROP2 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these spleen T cells. Values are expressed as mean ± SEM.
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Analysis of lymph node immune cells in B-hTROP2 mice
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Analysis of lymph node leukocyte subpopulations by flow cytometry. Leukocytes were isolated from female C57BL/6 and B-hTROP2 mice (n=3, 6 week-old). Flow cytometry analysis of the leukocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for CD45 population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells and NK cells in homozygous B-hTROP2 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hTROP2 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in lymph node. Values are expressed as mean ± SEM.
Analysis of lymph node T cell subpopulations by flow cytometry. Leukocytes were isolated from female C57BL/6 and B-hTROP2 mice (n=3, 6 week-old). Flow cytometry analysis of the leukocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for CD3 T cell population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of CD8+ T cells, CD4+ T cells and Treg cells in homozygous B-hTROP2 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hTROP2 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell sub types in lymph node. Values are expressed as mean ± SEM.
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Analysis of blood immune cells in B-hTROP2 mice
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Analysis of blood leukocyte subpopulations by flow cytometry. Blood cells were isolated from female C57BL/6 and B-hTROP2 mice (n=3, 6 week-old). Flow cytometry analysis of the blood leukocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for 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-hTROP2 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hTROP2 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in blood. Values are expressed as mean ± SEM.
Analysis of blood T cell subpopulations by flow cytometry. Blood cells were isolated from female C57BL/6 and B-hTROP2 mice (n=3, 6 week-old). Flow cytometry analysis of the leukocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for CD3 T cell population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of CD8+ T cells, CD4+ T cells and Treg cells in homozygous B-hTROP2 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hTROP2 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell sub types in blood. Values are expressed as mean ± SEM.
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Blood tests in B-hTROP2 mice
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Complete blood count (CBC). Blood was collected from wild-type C57BL/6 and B-hTROP2 mice (female, n=6, 9-week-old) and analyzed for CBC. Introduction of hTROP2 in place of its mouse counterpart does not change blood cell composition and morphology. Values are expressed as mean ± SEM.
Blood chemistry tests. Serum was collected from wild-type C57BL/6 and B-hTROP2 mice (n=6, 9-week-old) and analyzed for ALT and AST levels. Introduction of hTROP2 in place of its mouse counterpart does not change ALT and AST levels or liver health. Values are expressed as mean ± SEM.
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Poster
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Related mouse models
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