Analysis of MERTK gene expression in wild-type mice and B-hMERTK mice by q-PCR. The mRNA expression of MERTK in B-hMERTK (H/H) was similar to those in the wild-type mice (+/+), demonstrating that introduction of hMERTK in place of its mouse counterpart does not change the expression level of MERTK mRNA.  

Analysis of spleen leukocyte subpopulations by FACS. Splenocytes were isolated from male C57BL/6 and B-hMERTK mice(n=3, 7-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 NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hMERTK mice were similar to those in the C57BL/6 mice, demonstrating that B-hMERTK humanized does not change the overall development, differentiation or distribution of these cell types in spleen. Values are expressed as mean ± SEM.

Analysis of spleen T cell subpopulations by FACS. Splenocytes were isolated from male C57BL/6 and B-hMERTK mice(n=3, 7-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 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-hMERTK mice were similar to those in the C57BL/6 mice, demonstrating that introduction of B-hMERTK in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in spleen. Values are expressed as mean ± SEM.

Analysis of lymph node leukocyte subpopulations by FACS.Leukocytes were isolated from male C57BL/6 and B-hMERTK  mice(n=3, 7-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, NK cells, in homozygous B-hMERTK mice were similar to those in the C57BL/6 mice, demonstrating that introduction of B-hMERTK 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 leukocyte subpopulations by FACS. Leukocytes were isolated from male C57BL/6 and B-hMERTK mice(n=3, 7- week-old) Flow cytometry analysis of the leukocytes was performed to assess leukocyte subpopulations. Representative FACS plots. Single live CD45+ cells were gated for TCRb+ 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 in homozygous B-hMERTK mice were similar to those in the C57BL/6 mice,  demonstrating that introduction of B-hMERTK 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 blood leukocyte subpopulations by FACS. Leukocytes were isolated from male C57BL/6 and B-hMERTK mice(n=3, 7-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, NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hMERTK mice were similar to those in the C57BL/6 mice, demonstrating that introduction of B-hMERTKin 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 leukocyte subpopulations by FACS. Leukocytes were isolated from male C57BL/6 and B-hMERTKmice(n=3, 7-week-old) Flow cytometry analysis of the leukocytes was performed to assess leukocyte subpopulations. Representative FACS plots. Single live CD45+ cells were gated for TCRb+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD4+, CD8+, Tregs in homozygous B-hMERTK mice were similar to those in the C57BL/6 mice,  demonstrating that introduction of B-hMERTKin 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.