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R&D Systems sheep anti human osteolectin antibody
Sheep Anti Human Osteolectin Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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sheep anti human osteolectin antibody - by Bioz Stars, 2026-08

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( A, B ) The human ( A ) and mouse ( B ) Osteolectin proteins contain RGD and LDT sequences. ( C ) Alignment of Osteolectin amino acid sequences shows that the RGD and LDT domains are evolutionarily conserved among bony vertebrates. ( D, E ) RNA-seq analysis of integrin α ( D ) and β ( E ) subunits in PDGFRα + CD45 - Ter119 - CD31 - bone marrow stromal cells from enzymatically dissociated adult bone marrow (n = 2 independent samples). These cells are uniformly positive for LepR expression . ( F ) RNA-seq analysis of Itga1 , Itga6 , Itga11 , and Itgav in PDGFRα + CD45 - Ter119 - CD31 - bone marrow stromal cells, VE-Cadherin + bone marrow endothelial cells, and whole bone marrow cells (n = 2 independent samples per cell population). ( G ) Itga11 expression in cell populations from mouse bone marrow by qRT-PCR (n = 3 independent samples per cell population). The markers used for the isolation of each cell population are shown in . ( H ) In MC3T3-E1 preosteoblast cells expressing Flag-tagged Osteolectin, anti-Flag antibody co-immunoprecipitated endogenous integrin β1 and integrin α11 with Flag-tagged Osteolectin (results are representative of two independent experiments). ( I ) Recombinant human Osteolectin (rhOln) selectively bound to recombinant human integrin α 11 β 1 and α 10 β 1 , but not to other integrins (n = 3 independent experiments). ( J ) Integrin α11β1 bound Osteolectin and recombinant human Pro-Collagen 1α (rhCol1A) with similar affinities, but not bovine serum albumin (BSA) (n = 3 independent experiments). ( K ) Osteolectin, but not Pro-Collagen 1α, promoted osteogenic differentiation by MC3T3-E1 cells and human bone marrow stromal cells (n = 3 independent experiments). ( L ) 200 nM RGDS peptide inhibited the binding of integrin α11β1 to recombinant human Osteolectin. ( M ) 100 μM RGDS peptide inhibited osteogenic differentiation by MC3T3-E1 cells and human bone marrow stromal cells in response to 30 ng/ml of recombinant human Osteolectin. All numerical data reflect mean ±standard deviation. Statistical significance was determined with one-way ( G ) or two-way ANOVAs with Dunnett’s multiple comparisons tests ( K ) or Tukey’s multiple comparisons tests ( M ). 10.7554/eLife.42274.004 Figure 1—source data 1. Data for .

Journal: eLife

Article Title: Integrin alpha11 is an Osteolectin receptor and is required for the maintenance of adult skeletal bone mass

doi: 10.7554/eLife.42274

Figure Lengend Snippet: ( A, B ) The human ( A ) and mouse ( B ) Osteolectin proteins contain RGD and LDT sequences. ( C ) Alignment of Osteolectin amino acid sequences shows that the RGD and LDT domains are evolutionarily conserved among bony vertebrates. ( D, E ) RNA-seq analysis of integrin α ( D ) and β ( E ) subunits in PDGFRα + CD45 - Ter119 - CD31 - bone marrow stromal cells from enzymatically dissociated adult bone marrow (n = 2 independent samples). These cells are uniformly positive for LepR expression . ( F ) RNA-seq analysis of Itga1 , Itga6 , Itga11 , and Itgav in PDGFRα + CD45 - Ter119 - CD31 - bone marrow stromal cells, VE-Cadherin + bone marrow endothelial cells, and whole bone marrow cells (n = 2 independent samples per cell population). ( G ) Itga11 expression in cell populations from mouse bone marrow by qRT-PCR (n = 3 independent samples per cell population). The markers used for the isolation of each cell population are shown in . ( H ) In MC3T3-E1 preosteoblast cells expressing Flag-tagged Osteolectin, anti-Flag antibody co-immunoprecipitated endogenous integrin β1 and integrin α11 with Flag-tagged Osteolectin (results are representative of two independent experiments). ( I ) Recombinant human Osteolectin (rhOln) selectively bound to recombinant human integrin α 11 β 1 and α 10 β 1 , but not to other integrins (n = 3 independent experiments). ( J ) Integrin α11β1 bound Osteolectin and recombinant human Pro-Collagen 1α (rhCol1A) with similar affinities, but not bovine serum albumin (BSA) (n = 3 independent experiments). ( K ) Osteolectin, but not Pro-Collagen 1α, promoted osteogenic differentiation by MC3T3-E1 cells and human bone marrow stromal cells (n = 3 independent experiments). ( L ) 200 nM RGDS peptide inhibited the binding of integrin α11β1 to recombinant human Osteolectin. ( M ) 100 μM RGDS peptide inhibited osteogenic differentiation by MC3T3-E1 cells and human bone marrow stromal cells in response to 30 ng/ml of recombinant human Osteolectin. All numerical data reflect mean ±standard deviation. Statistical significance was determined with one-way ( G ) or two-way ANOVAs with Dunnett’s multiple comparisons tests ( K ) or Tukey’s multiple comparisons tests ( M ). 10.7554/eLife.42274.004 Figure 1—source data 1. Data for .

Article Snippet: Antibody , sheep polyclonal anti-human Osteolectin , R and D Systems , AF1904 , (1:1000).

Techniques: RNA Sequencing, Expressing, Quantitative RT-PCR, Isolation, Immunoprecipitation, Recombinant, Binding Assay, Standard Deviation

( A ) Western blot of cell culture supernatant from parental or Osteolectin deficient MC3T3-E1 cells and human bone marrow stromal cells (hBMSC#1 and hBMSC#2 cells) (this blot is representative of blots from three independent experiments). ( B ) Osteogenic differentiation in culture of parental or Osteolectin deficient MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells. Alizarin red staining was performed after 14 days (for MC3T3-E1 cells) or 21 days (for hBMSC cells) to quantify osteoblast differentiation and mineralization (n = 3 independent experiments). ( C ) qRT-PCR analysis of Dmp1 transcript levels in MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells after 14 to 21 days of osteogenic differentiation (n = 5 independent experiments). ( D ) MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells were stimulated with recombinant mouse Osteolectin (rOln) in osteogenic differentiation medium then lysed 0, 15, 30, or 60 min later and immunoblotted for phospho-PI3K, phospho-Akt, phospho-GSK3, β-catenin, total GSK3, and Actin (results are representative of 3 independent experiments). ( E ) MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells were transferred into osteogenic differentiation medium for 24 hr with PBS or recombinant mouse Osteolectin, lysed, and immunoblotted for phospho-GSK3, β-catenin, and Actin (this blot is representative of blots from three independent experiments). ( F ) qRT-PCR analysis of Wnt target gene transcript levels ( Alkaline phosphatase , Axin2 , Lef1 , or Runx2 ) in MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells 24 hr after transfer into osteogenic differentiation medium, with PBS or recombinant mouse Osteolectin (n = 5 independent experiments). ( G ) Parental or Osteolectin deficient MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells after 24 hr in osteogenic differentiation medium were lysed and immunoblotted for phospho-GSK3, β-catenin, and Actin (this blot is representative of blots from three independent experiments). ( H ) qRT-PCR analysis of Wnt target gene transcript levels in parental or Osteolectin deficient MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells 24 hr after transfer into osteogenic differentiation medium (n = 5 independent experiments). All numerical data reflect mean ±standard deviation. The statistical significance of differences was determined with two-way ANOVAs with Sidak’s multiple comparisons tests. 10.7554/eLife.42274.006 Figure 2—source data 1. Data for .

Journal: eLife

Article Title: Integrin alpha11 is an Osteolectin receptor and is required for the maintenance of adult skeletal bone mass

doi: 10.7554/eLife.42274

Figure Lengend Snippet: ( A ) Western blot of cell culture supernatant from parental or Osteolectin deficient MC3T3-E1 cells and human bone marrow stromal cells (hBMSC#1 and hBMSC#2 cells) (this blot is representative of blots from three independent experiments). ( B ) Osteogenic differentiation in culture of parental or Osteolectin deficient MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells. Alizarin red staining was performed after 14 days (for MC3T3-E1 cells) or 21 days (for hBMSC cells) to quantify osteoblast differentiation and mineralization (n = 3 independent experiments). ( C ) qRT-PCR analysis of Dmp1 transcript levels in MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells after 14 to 21 days of osteogenic differentiation (n = 5 independent experiments). ( D ) MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells were stimulated with recombinant mouse Osteolectin (rOln) in osteogenic differentiation medium then lysed 0, 15, 30, or 60 min later and immunoblotted for phospho-PI3K, phospho-Akt, phospho-GSK3, β-catenin, total GSK3, and Actin (results are representative of 3 independent experiments). ( E ) MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells were transferred into osteogenic differentiation medium for 24 hr with PBS or recombinant mouse Osteolectin, lysed, and immunoblotted for phospho-GSK3, β-catenin, and Actin (this blot is representative of blots from three independent experiments). ( F ) qRT-PCR analysis of Wnt target gene transcript levels ( Alkaline phosphatase , Axin2 , Lef1 , or Runx2 ) in MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells 24 hr after transfer into osteogenic differentiation medium, with PBS or recombinant mouse Osteolectin (n = 5 independent experiments). ( G ) Parental or Osteolectin deficient MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells after 24 hr in osteogenic differentiation medium were lysed and immunoblotted for phospho-GSK3, β-catenin, and Actin (this blot is representative of blots from three independent experiments). ( H ) qRT-PCR analysis of Wnt target gene transcript levels in parental or Osteolectin deficient MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells 24 hr after transfer into osteogenic differentiation medium (n = 5 independent experiments). All numerical data reflect mean ±standard deviation. The statistical significance of differences was determined with two-way ANOVAs with Sidak’s multiple comparisons tests. 10.7554/eLife.42274.006 Figure 2—source data 1. Data for .

Article Snippet: Antibody , sheep polyclonal anti-human Osteolectin , R and D Systems , AF1904 , (1:1000).

Techniques: Western Blot, Cell Culture, Staining, Quantitative RT-PCR, Recombinant, Standard Deviation

( A and B ) MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells were transferred into osteogenic differentiation medium with PBS or 30 ng/ml recombinant mouse Osteolectin, as well as DMSO or 200 nM of the GSK3 inhibitor AZD2858. ( A ) Cells were lysed 24 hr later and immunoblotted for phospho-GSK3, β-catenin, and Actin. ( B ) Alizarin red staining after 14 days (MC3T3-E1 cells) or 21 days (hBMSC cells) to quantify osteoblast differentiation and mineralization (n = 3 independent experiments). ( C and D ) MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells were transferred into osteogenic differentiation medium with PBS or 30 ng/ml Osteolectin, as well as DMSO or 200 nM of the β-catenin inhibitor IWR-1-endo. ( C ) Cells were lysed 24 hr later and immunoblotted for phospho-GSK3, β-catenin, and Actin. ( D ) Alizarin red staining after 14 days (MC3T3-E1 cells) or 21 days (hBMSC cells) to quantify osteoblast differentiation and mineralization (n = 3 independent experiments). All numerical data reflect mean ±standard deviation. The statistical significance of differences was determined with two-way ANOVAs with Tukey’s multiple comparisons tests. 10.7554/eLife.42274.008 Figure 3—source data 1. Data for Data for .

Journal: eLife

Article Title: Integrin alpha11 is an Osteolectin receptor and is required for the maintenance of adult skeletal bone mass

doi: 10.7554/eLife.42274

Figure Lengend Snippet: ( A and B ) MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells were transferred into osteogenic differentiation medium with PBS or 30 ng/ml recombinant mouse Osteolectin, as well as DMSO or 200 nM of the GSK3 inhibitor AZD2858. ( A ) Cells were lysed 24 hr later and immunoblotted for phospho-GSK3, β-catenin, and Actin. ( B ) Alizarin red staining after 14 days (MC3T3-E1 cells) or 21 days (hBMSC cells) to quantify osteoblast differentiation and mineralization (n = 3 independent experiments). ( C and D ) MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells were transferred into osteogenic differentiation medium with PBS or 30 ng/ml Osteolectin, as well as DMSO or 200 nM of the β-catenin inhibitor IWR-1-endo. ( C ) Cells were lysed 24 hr later and immunoblotted for phospho-GSK3, β-catenin, and Actin. ( D ) Alizarin red staining after 14 days (MC3T3-E1 cells) or 21 days (hBMSC cells) to quantify osteoblast differentiation and mineralization (n = 3 independent experiments). All numerical data reflect mean ±standard deviation. The statistical significance of differences was determined with two-way ANOVAs with Tukey’s multiple comparisons tests. 10.7554/eLife.42274.008 Figure 3—source data 1. Data for Data for .

Article Snippet: Antibody , sheep polyclonal anti-human Osteolectin , R and D Systems , AF1904 , (1:1000).

Techniques: Recombinant, Staining, Standard Deviation

$( A ) Osteogenic differentiation in culture of parental or Itga11 deficient MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells with PBS or recombinant mouse Osteolectin (n = 3 independent experiments). ( B ) qRT-PCR analysis of Dmp1 transcript levels in MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells after 14 to 21 days of osteogenic differentiation (n = 5 independent experiments). ( C ) Parental or Itga11 deficient MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells were transferred into osteogenic differentiation medium with or without recombinant mouse Osteolectin then lysed 24 hr later and immunoblotted for phospho-GSK3, β-catenin, and Actin (this blot is representative of blots from three independent experiments). ( D ) qRT-PCR analysis of Wnt target gene transcript levels in parental or Itga11 deficient MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells 24 hr after transfer into osteogenic differentiation medium with PBS or Osteolectin (n = 5 independent experiments). ( E ) MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells were transferred into osteogenic differentiation medium with PBS or 30 ng/ml Osteolectin, or 30 ng/ml recombinant Pro-Collagen 1α, then lysed 24 hr later and immunoblotted for phospho-GSK3, β-catenin, and Actin (this blot is representative of blots from three independent experiments). ( F ) qRT-PCR analysis of Wnt target gene transcript levels in MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells 24 hr after transfer into osteogenic differentiation medium with PBS or 30 ng/ml Osteolectin, or 30 ng/ml Pro-Collagen 1α (n = 5 independent experiments). ( G ) Primary mouse bone marrow stromal cells were adherently cultured in osteogenic differentiation medium. PBS (control), recombinant human Pro-Collagen 1α, or recombinant human Osteolectin was added, then the cells were lysed 1 hr later and lysates were immunoblotted for phospho-FAK, phospho-GSK3, total FAK, and Actin (this blot is representative of blots from three independent experiments). ( H ) Primary mouse bone marrow stromal cells adherently cultured in osteogenic differentiation medium were treated with PBS (control), recombinant human Pro-Collagen 1α, or recombinant human Osteolectin then lysed 24 hr later and lysates were immunoblotted for phospho-FAK, phospho-GSK3, β-catenin, total FAK, and Actin (this blot is representative of blots from three independent experiments). ( I ) Primary mouse bone marrow stromal cells adherently cultured in osteogenic differentiation medium were treated with 30 ng/ml recombinant human Osteolectin or PBS (control) then lysed 2, 4, 6, 12, or 24 hr later. Nuclear and cytosolic/membrane-associated fractions were isolated from lysates by centrifugation then immunoblotted for β-catenin. As loading controls, Histone H3 was blotted in the nuclear fraction and Actin was blotted in the cytosolic/membrane-associated fraction (this blot is representative of blots from three independent experiments). ( J ) qRT-PCR analysis of Ctnnb1 transcript levels in cells from the experiment in panel ( I ) (n = 3 independent experiments). ( K ) Primary mouse bone marrow stromal cells from Lepr-Cre; Itga11 fl/fl or littermate control mice were adherently cultured in osteogenic differentiation medium. PBS (control), recombinant human Pro-Collagen 1α, or recombinant human Osteolectin was added then the cells were lysed 1 hr later and lysates were immunoblotted for phospho-FAK and FAK (this blot is representative of blots from two independent experiments). All numerical data reflect mean ±standard deviation. Statistical significance was determined with two-way ANOVAs with Tukey’s multiple comparisons tests ( A, B and D ), Dunnett’s multiple comparisons tests ( F ), or Sidak’s multiple comparisons tests ( J ). 10.7554/eLife.42274.010 Figure 4—source data 1. Data for .$

Journal: eLife

Article Title: Integrin alpha11 is an Osteolectin receptor and is required for the maintenance of adult skeletal bone mass

doi: 10.7554/eLife.42274

Figure Lengend Snippet: ( A ) Osteogenic differentiation in culture of parental or Itga11 deficient MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells with PBS or recombinant mouse Osteolectin (n = 3 independent experiments). ( B ) qRT-PCR analysis of Dmp1 transcript levels in MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells after 14 to 21 days of osteogenic differentiation (n = 5 independent experiments). ( C ) Parental or Itga11 deficient MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells were transferred into osteogenic differentiation medium with or without recombinant mouse Osteolectin then lysed 24 hr later and immunoblotted for phospho-GSK3, β-catenin, and Actin (this blot is representative of blots from three independent experiments). ( D ) qRT-PCR analysis of Wnt target gene transcript levels in parental or Itga11 deficient MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells 24 hr after transfer into osteogenic differentiation medium with PBS or Osteolectin (n = 5 independent experiments). ( E ) MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells were transferred into osteogenic differentiation medium with PBS or 30 ng/ml Osteolectin, or 30 ng/ml recombinant Pro-Collagen 1α, then lysed 24 hr later and immunoblotted for phospho-GSK3, β-catenin, and Actin (this blot is representative of blots from three independent experiments). ( F ) qRT-PCR analysis of Wnt target gene transcript levels in MC3T3-E1 cells, hBMSC#1 cells, and hBMSC#2 cells 24 hr after transfer into osteogenic differentiation medium with PBS or 30 ng/ml Osteolectin, or 30 ng/ml Pro-Collagen 1α (n = 5 independent experiments). ( G ) Primary mouse bone marrow stromal cells were adherently cultured in osteogenic differentiation medium. PBS (control), recombinant human Pro-Collagen 1α, or recombinant human Osteolectin was added, then the cells were lysed 1 hr later and lysates were immunoblotted for phospho-FAK, phospho-GSK3, total FAK, and Actin (this blot is representative of blots from three independent experiments). ( H ) Primary mouse bone marrow stromal cells adherently cultured in osteogenic differentiation medium were treated with PBS (control), recombinant human Pro-Collagen 1α, or recombinant human Osteolectin then lysed 24 hr later and lysates were immunoblotted for phospho-FAK, phospho-GSK3, β-catenin, total FAK, and Actin (this blot is representative of blots from three independent experiments). ( I ) Primary mouse bone marrow stromal cells adherently cultured in osteogenic differentiation medium were treated with 30 ng/ml recombinant human Osteolectin or PBS (control) then lysed 2, 4, 6, 12, or 24 hr later. Nuclear and cytosolic/membrane-associated fractions were isolated from lysates by centrifugation then immunoblotted for β-catenin. As loading controls, Histone H3 was blotted in the nuclear fraction and Actin was blotted in the cytosolic/membrane-associated fraction (this blot is representative of blots from three independent experiments). ( J ) qRT-PCR analysis of Ctnnb1 transcript levels in cells from the experiment in panel ( I ) (n = 3 independent experiments). ( K ) Primary mouse bone marrow stromal cells from Lepr-Cre; Itga11 fl/fl or littermate control mice were adherently cultured in osteogenic differentiation medium. PBS (control), recombinant human Pro-Collagen 1α, or recombinant human Osteolectin was added then the cells were lysed 1 hr later and lysates were immunoblotted for phospho-FAK and FAK (this blot is representative of blots from two independent experiments). All numerical data reflect mean ±standard deviation. Statistical significance was determined with two-way ANOVAs with Tukey’s multiple comparisons tests ( A, B and D ), Dunnett’s multiple comparisons tests ( F ), or Sidak’s multiple comparisons tests ( J ). 10.7554/eLife.42274.010 Figure 4—source data 1. Data for .

Article Snippet: Antibody , sheep polyclonal anti-human Osteolectin , R and D Systems , AF1904 , (1:1000).

Techniques: Recombinant, Quantitative RT-PCR, Cell Culture, Control, Membrane, Isolation, Centrifugation, Standard Deviation

( A ) Lepr-Cre; Itga11 fl/fl mice were grossly normal and indistinguishable from littermate controls. ( B – D ) Body length ( B ), body mass ( C ) and femur length ( D ) did not significantly differ between Lepr-Cre; Itga11 fl/fl mice and sex-matched littermate controls at 2, 6, or 12 months of age (n = 4–7 mice per genotype per sex per time point, from at least three independent experiments). ( E ) ELISA measurement of serum Osteolectin levels in Lepr-Cre; Itga11 fl/fl mice and littermate controls at 2, 6, and 12 months of age (n = 8 mice per genotype per time point from four independent experiments). ( F ) Representative microCT images of trabecular bone in the distal femur metaphysis of male Lepr-Cre; Itga11 fl/fl mice and littermate controls at 2, 6, and 12 months of age. ( G–L ) microCT analysis of trabecular bone volume/total volume ( G ), trabecular number ( H ), trabecular bone thickness ( I ), trabecular bone spacing ( J ), connectivity density ( K ), and bone mineral density ( L ) in the distal femur metaphysis of Lepr-Cre; Itga11 fl/fl mice and sex-matched littermate controls at 2, 6, and 12 months of age (n = 4–7 mice per genotype per sex per time point from at least three independent experiments). ( M ) Trabecular bone mineral apposition rate based on calcein double labelling in the distal femur metaphysis (n = 3 mice per genotype per sex per time point). ( N ) Bone resorption rate analysis by measuring the deoxypyridinoline/creatinine ratio in the urine of LepR-Cre; Itga11 fl/fl mice and littermate controls at 6 and 12 months of age (n = 4–5 mice per genotype per time point from three independent experiments). ( O ) Serum P1NP levels in LepR-Cre; Itga11 fl/fl mice and sex-matched littermate controls at 6 and 12 months of age (n = 4 mice per genotype per time point from three independent experiments). All numerical data reflect mean ±standard deviation. The statistical significance of differences was determined with two-way ANOVAs with Sidak’s multiple comparisons tests. 10.7554/eLife.42274.012 Figure 5—source data 1. Data for Data for .

Journal: eLife

Article Title: Integrin alpha11 is an Osteolectin receptor and is required for the maintenance of adult skeletal bone mass

doi: 10.7554/eLife.42274

Figure Lengend Snippet: ( A ) Lepr-Cre; Itga11 fl/fl mice were grossly normal and indistinguishable from littermate controls. ( B – D ) Body length ( B ), body mass ( C ) and femur length ( D ) did not significantly differ between Lepr-Cre; Itga11 fl/fl mice and sex-matched littermate controls at 2, 6, or 12 months of age (n = 4–7 mice per genotype per sex per time point, from at least three independent experiments). ( E ) ELISA measurement of serum Osteolectin levels in Lepr-Cre; Itga11 fl/fl mice and littermate controls at 2, 6, and 12 months of age (n = 8 mice per genotype per time point from four independent experiments). ( F ) Representative microCT images of trabecular bone in the distal femur metaphysis of male Lepr-Cre; Itga11 fl/fl mice and littermate controls at 2, 6, and 12 months of age. ( G–L ) microCT analysis of trabecular bone volume/total volume ( G ), trabecular number ( H ), trabecular bone thickness ( I ), trabecular bone spacing ( J ), connectivity density ( K ), and bone mineral density ( L ) in the distal femur metaphysis of Lepr-Cre; Itga11 fl/fl mice and sex-matched littermate controls at 2, 6, and 12 months of age (n = 4–7 mice per genotype per sex per time point from at least three independent experiments). ( M ) Trabecular bone mineral apposition rate based on calcein double labelling in the distal femur metaphysis (n = 3 mice per genotype per sex per time point). ( N ) Bone resorption rate analysis by measuring the deoxypyridinoline/creatinine ratio in the urine of LepR-Cre; Itga11 fl/fl mice and littermate controls at 6 and 12 months of age (n = 4–5 mice per genotype per time point from three independent experiments). ( O ) Serum P1NP levels in LepR-Cre; Itga11 fl/fl mice and sex-matched littermate controls at 6 and 12 months of age (n = 4 mice per genotype per time point from three independent experiments). All numerical data reflect mean ±standard deviation. The statistical significance of differences was determined with two-way ANOVAs with Sidak’s multiple comparisons tests. 10.7554/eLife.42274.012 Figure 5—source data 1. Data for Data for .

Article Snippet: Antibody , sheep polyclonal anti-human Osteolectin , R and D Systems , AF1904 , (1:1000).

Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation

( A ) CFU-F frequency and ( B ) cells per CFU-F colony formed by bone marrow cells from Lepr-Cre; Itga11 fl/fl mice and littermate controls at 2 and 6 months of age (n = 6–8 mice per genotype per time point from at least three independent experiments). ( C–E ) Osteogenic ( C ); n = 7 mice per genotype, total, from seven independent experiments), adipogenic ( D ); n = 6 mice per genotype, total, from six independent experiments), and chondrogenic ( E ); n = 5 mice per genotype, total, from five independent experiments) differentiation of bone marrow stromal cells cultured from the femurs of Lepr-Cre; Itga11 fl/fl mice and sex-matched littermate controls at 2 months of age. ( F ) Recombinant mouse Osteolectin promoted osteogenic differentiation in culture by femur bone marrow stromal cells from control but not Lepr-Cre; Itga11 fl/fl mice (n = 3 mice per genotype, total, from three independent experiments). ( G ) qRT-PCR analysis of Dmp1 transcript levels in cells from panel ( F ). ( H–N ) Subcutaneous injection of recombinant mouse Osteolectin daily for 28 days (50 μg/kg body mass/day) significantly increased trabecular bone volume and number in female control mice but not female Lepr-Cre; Itga11 fl/fl mice. ( H ) Representative microCT images of trabecular bone in the distal femur metaphysis. ( I–N ) microCT analysis of trabecular bone volume/total volume ( I ), trabecular number ( J ), trabecular thickness ( K ), trabecular spacing ( L ), connectivity density ( M ), and bone mineral density ( N ) in the distal femur metaphysis (n = 5 mice per treatment, total, from three independent experiments). ( O ) The GSK3 inhibitor, AZD2858, rescued the osteogenic differentiation in culture of bone marrow stromal cells from the femurs of Lepr-Cre; Itga11 fl/fl mice (n = 6 independent experiments in which cells from one mouse of each genotype were cultured in each experiment). ( P ) Western blotting of cultured cell lysates showed that AZD2858 promoted GSK3 phosphorylation and increased β-catenin levels in bone marrow stromal cells from Lepr-Cre; Itga11 fl/fl mice and littermate controls. All numerical data reflect mean ±standard deviation. The statistical significance of differences was determined with Wilcoxon’s test followed by Holm-Sidak multiple comparisons adjustment ( A and B ), one-way ( F ), ( G, ) and O ) ANOVAs with Sidak’s multiple comparisons tests, by paired t-tests ( C–E ), or by one-way ANOVAs with Tukey’s multiple comparisons tests ( I–N ). 10.7554/eLife.42274.018 Figure 7—source data 1. Data for .

Journal: eLife

Article Title: Integrin alpha11 is an Osteolectin receptor and is required for the maintenance of adult skeletal bone mass

doi: 10.7554/eLife.42274

Figure Lengend Snippet: ( A ) CFU-F frequency and ( B ) cells per CFU-F colony formed by bone marrow cells from Lepr-Cre; Itga11 fl/fl mice and littermate controls at 2 and 6 months of age (n = 6–8 mice per genotype per time point from at least three independent experiments). ( C–E ) Osteogenic ( C ); n = 7 mice per genotype, total, from seven independent experiments), adipogenic ( D ); n = 6 mice per genotype, total, from six independent experiments), and chondrogenic ( E ); n = 5 mice per genotype, total, from five independent experiments) differentiation of bone marrow stromal cells cultured from the femurs of Lepr-Cre; Itga11 fl/fl mice and sex-matched littermate controls at 2 months of age. ( F ) Recombinant mouse Osteolectin promoted osteogenic differentiation in culture by femur bone marrow stromal cells from control but not Lepr-Cre; Itga11 fl/fl mice (n = 3 mice per genotype, total, from three independent experiments). ( G ) qRT-PCR analysis of Dmp1 transcript levels in cells from panel ( F ). ( H–N ) Subcutaneous injection of recombinant mouse Osteolectin daily for 28 days (50 μg/kg body mass/day) significantly increased trabecular bone volume and number in female control mice but not female Lepr-Cre; Itga11 fl/fl mice. ( H ) Representative microCT images of trabecular bone in the distal femur metaphysis. ( I–N ) microCT analysis of trabecular bone volume/total volume ( I ), trabecular number ( J ), trabecular thickness ( K ), trabecular spacing ( L ), connectivity density ( M ), and bone mineral density ( N ) in the distal femur metaphysis (n = 5 mice per treatment, total, from three independent experiments). ( O ) The GSK3 inhibitor, AZD2858, rescued the osteogenic differentiation in culture of bone marrow stromal cells from the femurs of Lepr-Cre; Itga11 fl/fl mice (n = 6 independent experiments in which cells from one mouse of each genotype were cultured in each experiment). ( P ) Western blotting of cultured cell lysates showed that AZD2858 promoted GSK3 phosphorylation and increased β-catenin levels in bone marrow stromal cells from Lepr-Cre; Itga11 fl/fl mice and littermate controls. All numerical data reflect mean ±standard deviation. The statistical significance of differences was determined with Wilcoxon’s test followed by Holm-Sidak multiple comparisons adjustment ( A and B ), one-way ( F ), ( G, ) and O ) ANOVAs with Sidak’s multiple comparisons tests, by paired t-tests ( C–E ), or by one-way ANOVAs with Tukey’s multiple comparisons tests ( I–N ). 10.7554/eLife.42274.018 Figure 7—source data 1. Data for .

Article Snippet: Antibody , sheep polyclonal anti-human Osteolectin , R and D Systems , AF1904 , (1:1000).

Techniques: Cell Culture, Recombinant, Control, Quantitative RT-PCR, Injection, Western Blot, Phospho-proteomics, Standard Deviation

( A ) Bone marrow stromal cells from Lepr-Cre; Itga11 fl/fl mice and sex-matched littermates at 2 months of age were cultured in osteogenic differentiation medium supplemented with PBS (control) or 30 ng/ml Osteolectin and qRT-PCR analysis of Wnt target gene transcript levels was performed 24 hr later (n = 3 independent experiments). ( B ) Recombinant mouse Osteolectin subcutaneously injected daily for 28 days (50 μg/kg body mass/day) increased Wnt target gene transcript levels in LepR + bone marrow cells from control, but not Lepr-Cre; Itga11 fl/fl , mice (n = 4 mice per genotype per time point from two independent experiments). All numerical data reflect mean ±standard deviation. Statistical significance was assessed with two-way ANOVAs with Tukey’s multiple comparisons tests ( A ) or Dunnett’s multiple comparisons tests ( B ). 10.7554/eLife.42274.020 Figure 7—figure supplement 1—source data 1. Data for .

Journal: eLife

Article Title: Integrin alpha11 is an Osteolectin receptor and is required for the maintenance of adult skeletal bone mass

doi: 10.7554/eLife.42274

Figure Lengend Snippet: ( A ) Bone marrow stromal cells from Lepr-Cre; Itga11 fl/fl mice and sex-matched littermates at 2 months of age were cultured in osteogenic differentiation medium supplemented with PBS (control) or 30 ng/ml Osteolectin and qRT-PCR analysis of Wnt target gene transcript levels was performed 24 hr later (n = 3 independent experiments). ( B ) Recombinant mouse Osteolectin subcutaneously injected daily for 28 days (50 μg/kg body mass/day) increased Wnt target gene transcript levels in LepR + bone marrow cells from control, but not Lepr-Cre; Itga11 fl/fl , mice (n = 4 mice per genotype per time point from two independent experiments). All numerical data reflect mean ±standard deviation. Statistical significance was assessed with two-way ANOVAs with Tukey’s multiple comparisons tests ( A ) or Dunnett’s multiple comparisons tests ( B ). 10.7554/eLife.42274.020 Figure 7—figure supplement 1—source data 1. Data for .

Article Snippet: Antibody , sheep polyclonal anti-human Osteolectin , R and D Systems , AF1904 , (1:1000).

Techniques: Cell Culture, Control, Quantitative RT-PCR, Recombinant, Injection, Standard Deviation

Journal: eLife

Article Title: Integrin alpha11 is an Osteolectin receptor and is required for the maintenance of adult skeletal bone mass

doi: 10.7554/eLife.42274

Figure Lengend Snippet:

Article Snippet: Antibody , sheep polyclonal anti-human Osteolectin , R and D Systems , AF1904 , (1:1000).

Techniques: Recombinant, Diagnostic Assay, Cell Culture, Protease Inhibitor, Western Blot, Reverse Transcription, Enzyme-linked Immunosorbent Assay, Fractionation

$RNA-seq analysis of LepR + cells from non-irradiated bone marrow as well as hematopoietic cells and unfractionated cells from irradiated bone marrow (which are high in adipocytes). Itga11 is poorly expressed in adipocyte-rich bone marrow cells after irradiation (all data represent average FPKM values from multiple replicates).$

Journal: eLife

Article Title: Integrin alpha11 is an Osteolectin receptor and is required for the maintenance of adult skeletal bone mass

doi: 10.7554/eLife.42274

Figure Lengend Snippet: RNA-seq analysis of LepR + cells from non-irradiated bone marrow as well as hematopoietic cells and unfractionated cells from irradiated bone marrow (which are high in adipocytes). Itga11 is poorly expressed in adipocyte-rich bone marrow cells after irradiation (all data represent average FPKM values from multiple replicates).

Article Snippet: Antibody , sheep polyclonal anti-human Osteolectin , R and D Systems , AF1904 , (1:1000).

Techniques: Irradiation

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