请输入关键字
请输入关键字
订购
*国家
中国
美国
*省份
*城市
*姓名
*电话
*单位
*职位
*邮箱
*请输入验证码
*验证码
B-hRANKL mice
Strain Name C57BL/6-Tnfsf11tm1(TNFSF11)/Bcgen Common Name  B-hRANKL mice
Background C57BL/6 Catalog number  111072
Aliases 
CD254, ODF, OPGL, OPTB2, RANKL, TNLG6B, TRANCE, hRANKL2, sOdf

Gene Description


RANKL encodes a member of the tumor necrosis factor (TNF) cytokine family which is a ligand for osteoprotegerin and functions as a key factor for osteoclast differentiation and activation. This protein was shown to be a dendritic cell survival factor and is involved in the regulation of T cell-dependent immune response. T cell activation was reported to induce expression of this gene and lead to an increase of osteoclastogenesis and bone loss. This protein was shown to activate antiapoptotic kinase AKT/PKB through a signaling complex involving SRC kinase and tumor necrosis factor receptor-associated factor (TRAF) 6, which indicated this protein may have a role in the regulation of cell apoptosis. Targeted disruption of the related gene in mice led to severe osteopetrosis and a lack of osteoclasts. The deficient mice exhibited defects in early differentiation of T and B lymphocytes, and failed to form lobulo-alveolar mammary structures during pregnancy. 


Protein expression analysis


from clipboard


Strain specific RANKL expression analysis in wild-type C57BL/6 mice and homozygous B-hRANKL mice by flow cytometry. CD4+ T cells were isolated from the spleen of wild-type C57BL/6 mice (+/+) and homozygous B-hRANKL mice (H/H) stimulated with anti-mCD3e and anti-mCD28 antibodies in vitro. Mouse RANKL was detectable only in wild-type C57BL/6 mice. Human RANKL was exclusively detectable in homozygous B-hRANKL mice.


mRNA expression analysis



Strain specific analysis of RANKL gene expression in wild-type C57BL/6 mice and B-hRANKL mice by RT-PCR. Mouse Rankl mRNA was detectable only in thymocytes of wild-type C57BL/6 mice (+/+). Human RANKL mRNA was detectable only in homozygous B-hRANKL mice but not in wild-type mice. 


Analysis of leukocytes cell subpopulation in spleen


from clipboard


Analysis of spleen leukocyte subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hRANKL 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-hRANKL mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hRANKL 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 T cell subpopulation in spleen

from clipboard


Analysis of spleen T cell subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hRANKL 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 CD4+ T cells, CD8+ T cells and Tregs in homozygous B-hRANKL mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hRANKL 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 leukocytes cell subpopulation in lymph node

from clipboard


Analysis of lymph node leukocyte subpopulations by FACS. Lymph nodes were isolated from female C57BL/6 and B-hRANKL 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 the 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-hRANKL mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hRANKL in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in lymph node. Values are expressed as mean ± SEM.


Analysis of T cell subpopulation in lymph node

from clipboard


Analysis of lymph node T cell subpopulations by FACS. Leukocytes were isolated from female C57BL/6 and B-hRANKL 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 TCRβ+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD4+ T cells, CD8+ T cells, and Tregs in homozygous B-hRANKL mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hRANKL in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in lymph node. Values are expressed as mean ± SEM.


Analysis of leukocytes cell subpopulation in blood

from clipboard


Analysis of blood leukocyte subpopulations by FACS. Blood cells were isolated from female C57BL/6 and B-hRANKL mice (n=3, 6-week-old). Flow cytometry analysis of the blood cells 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-hRANKL mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hRANKL in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in blood. Values are expressed as mean ± SEM.


Analysis of T cell subpopulation in blood

from clipboard


Analysis of blood T cell subpopulations by FACS. Blood cells were isolated from female C57BL/6 and B-hRANKL mice (n=3, 6-week-old). Flow cytometry analysis of the blood cells 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 CD4+ T cells, CD8+ T cells, and Tregs in homozygous B-hRANKL mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hRANKL in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in blood. Values are expressed as mean ± SEM.