human rankl Search Results


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  • 98
    BioVendor Instruments soluble rankl
    Soluble Rankl, supplied by BioVendor Instruments, used in various techniques. Bioz Stars score: 98/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher human recombinant rankl
    Human Recombinant Rankl, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Amgen human rankl
    A : Immunohistochemical analysis of <t>RANKL</t> in <t>C4-2</t> tumored tibiae of control, prevention, and treatment groups. Paraffin-embedded C4-2 tumored tibiae of the SCID mice were sectioned and stained for RANKL (negative control staining is shown in inset box). The tumor cells stained positively for RANKL. B : Blood was collected to determine serum PSA levels by IMx Total PSA assay. Serum PSA levels were not significantly different between the C4-2 control with PBS injections (control), C4-2 with a concomitant subcutaneous injection of huRANKL MAb (5 mg/kg once a week) at implantation (prevention group), or C4-2 with a subcutaneous injection of huRANKL MAb (5 mg/kg biweekly, starting 3 weeks after implantation of C4-2 cells) (treatment group). Results are presented as mean ± SE.
    Human Rankl, supplied by Amgen, used in various techniques. Bioz Stars score: 90/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    R&D Systems recombinant human rankl
    S100A12 stimulates the expression of mature-osteoclast markers. Human CD14 + monocytes were cultured in the presence of <t>M-CSF</t> (50 ng/ml), alone or in combination with <t>RANKL</t> (100 ng/ml) and with or without S100A12 (1,000 ng/ml) for 7–9 days. Levels of mRNA expression for nuclear factor of activated T cells c1 (NF-ATc1), TRAP (ACP5), calcitonin receptor (CALCR), integrin β3 (ITGβ3), cathepsin K (CTSK), and carbonic anhydrase II (CA2) were determined by qRT-PCR. Messenger RNA levels were normalized to GAPDH. Values are the mean and SD of 7 independent experiments. * = p
    Recombinant Human Rankl, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 61 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore recombinant human rankl
    The effect of Hmox1 knockout in osteoclasts precursors on OCLs-specific genes expression. Bone marrow was isolated from HO-1 −/− and HO-1 +/+ mice. ( A , B ) BMCs-derived BMMs were stimulated with <t>RANKL</t> (50 ng/ml or 100 ng/ml) in the presence of 100 ng/ml <t>M-CSF</t> for 3 days. Alternatively, ( C , D ) BMCs-derived BMMs and ( E , F ) nBMCs-derived BMMs were replated and cultured in the presence of 50 ng/ml RANKL and 30 ng/ml M-CSF for 3 days. ( A , C , E ) NFAT-c1 and ( B , D , F ) cathepsin K relative expression (vs. EF-2). Quantitative PCR (n A,B = 9–10, n C,D = 7, n E,F = 17–20). Each bar represents the mean ± SEM. *p
    Recombinant Human Rankl, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    R&D Systems human trance tnfsf11 rank l antibody
    The effect of Hmox1 knockout in osteoclasts precursors on OCLs-specific genes expression. Bone marrow was isolated from HO-1 −/− and HO-1 +/+ mice. ( A , B ) BMCs-derived BMMs were stimulated with <t>RANKL</t> (50 ng/ml or 100 ng/ml) in the presence of 100 ng/ml <t>M-CSF</t> for 3 days. Alternatively, ( C , D ) BMCs-derived BMMs and ( E , F ) nBMCs-derived BMMs were replated and cultured in the presence of 50 ng/ml RANKL and 30 ng/ml M-CSF for 3 days. ( A , C , E ) NFAT-c1 and ( B , D , F ) cathepsin K relative expression (vs. EF-2). Quantitative PCR (n A,B = 9–10, n C,D = 7, n E,F = 17–20). Each bar represents the mean ± SEM. *p
    Human Trance Tnfsf11 Rank L Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 91/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    PeproTech rankl
    The effect of Hmox1 knockout in osteoclasts precursors on OCLs-specific genes expression. Bone marrow was isolated from HO-1 −/− and HO-1 +/+ mice. ( A , B ) BMCs-derived BMMs were stimulated with <t>RANKL</t> (50 ng/ml or 100 ng/ml) in the presence of 100 ng/ml <t>M-CSF</t> for 3 days. Alternatively, ( C , D ) BMCs-derived BMMs and ( E , F ) nBMCs-derived BMMs were replated and cultured in the presence of 50 ng/ml RANKL and 30 ng/ml M-CSF for 3 days. ( A , C , E ) NFAT-c1 and ( B , D , F ) cathepsin K relative expression (vs. EF-2). Quantitative PCR (n A,B = 9–10, n C,D = 7, n E,F = 17–20). Each bar represents the mean ± SEM. *p
    Rankl, supplied by PeproTech, used in various techniques. Bioz Stars score: 94/100, based on 1538 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems recombinant human trance rank l tnfsf11 protein
    The effect of Hmox1 knockout in osteoclasts precursors on OCLs-specific genes expression. Bone marrow was isolated from HO-1 −/− and HO-1 +/+ mice. ( A , B ) BMCs-derived BMMs were stimulated with <t>RANKL</t> (50 ng/ml or 100 ng/ml) in the presence of 100 ng/ml <t>M-CSF</t> for 3 days. Alternatively, ( C , D ) BMCs-derived BMMs and ( E , F ) nBMCs-derived BMMs were replated and cultured in the presence of 50 ng/ml RANKL and 30 ng/ml M-CSF for 3 days. ( A , C , E ) NFAT-c1 and ( B , D , F ) cathepsin K relative expression (vs. EF-2). Quantitative PCR (n A,B = 9–10, n C,D = 7, n E,F = 17–20). Each bar represents the mean ± SEM. *p
    Recombinant Human Trance Rank L Tnfsf11 Protein, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PeproTech recombinant human soluble rankl
    Microscopic image of a cultured osteoclast. Murine bone marrow macrophages were incubated for 82 h with <t>RANKL</t> (100 ng/mL) and <t>M-CSF</t> (10 ng/mL), resulting in osteoclast formation. After culturing, cells were fixed with 4% paraformaldehyde in phosphate buffer and stained with DAPI and Rhodamine B-conjugated phalloidin. Osteoclasts are multinucleated giant cells with a ring-shaped osteoclast-specific podosome belt termed the actin ring. Blue, nuclei; red, actin; scale bar = 50 µm.
    Recombinant Human Soluble Rankl, supplied by PeproTech, used in various techniques. Bioz Stars score: 99/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PeproTech recombinant human rankl
    Inhibition of osteoclastogenesis by ASMSCs compared with HDMSCs. CD14 + monocytes were cultured with HDMSCs or ASMSCs in the presence of <t>M-CSF</t> and <t>RANKL.</t> a Representative images of TRAP staining of osteoclasts co-cultured at different time points (× 100). b The number of TRAP + osteoclasts in each well from cultures at different time points is shown. c Representative images of osteoclasts stained with FITC-phalloidin at different time points (× 200). d Representative images for bone resorption assays at different time points ( × 200). Cells cultured with HDMSCs or ASMSCs on bovine cortical slides were stained with toluidine blue. e Pit formation on each slide was assessed. f mRNA expression levels of TRAP, CTSK, and NFATc1 in osteoclasts were determined by qPCR on day 9. g Protein levels of TRAP, CTSK, and NFATc1 in osteoclasts were determined by western blot analyses on day 9. h Quantitative data of TRAP, CTSK, and NFATc1 protein levels determined by western blot analyses are shown. i Activation of signaling pathways involved in osteoclastogenesis was determined by western blot analyses on day 9. j Quantitative data for activation of signaling pathways determined by western blot analyses are shown. Values are the mean ± SD of 30 samples per group. The results represent three independent experiments. *, p
    Recombinant Human Rankl, supplied by PeproTech, used in various techniques. Bioz Stars score: 94/100, based on 109 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Santa Cruz Biotechnology human rankl
    Effects of NGF upon osteoclast differentiation in vitro . Human peripheral mononuclear cells were isolated and cultured in the presence of <t>MCSF</t> with (A) or without (B) <t>RANKL,</t> and increasing concentrations of NGF, for 14 days. TRAP +ve cells were counted using four random fields of vision on four separate slides for each condition. RANKL alone significantly increased TRAP +ve cells as compared to MCSF alone (A). For RANKL treated cells, addition of NGF significantly decreased osteoclast numbers at 50 and 100 ng/ml. In the absence of RANKL (B), addition of NGF caused a small non-dose related significant increase in TRAP +ve cell number. (C) Examples of TRAP staining in the presence and absence of RANKL. Scale bar is 100 μm. Data are mean ± SEM, statistical analysis: one way ANOVA, * P
    Human Rankl, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Abcam anti human rankl antibody
    Methotrexate decreases expression of <t>RANKL</t> and <t>RANK</t> in the rheumatoid synovium of early-untreated rheumatoid arthritis. Brown immunohistochemistry (diaminobenzidine) staining of cell surface markers in synovial biopsies of early rheumatoid arthritis (RA). Synovial receptor activator of the NF-κB ligand (RANKL) expression in early, untreated RA synovium (A) decreased significantly following 8 weeks of methotrexate (MTX) treatment (B) , as evaluated by semi-quantitative double-blind microscopic analysis (C) . Synovial receptor activator of NF-κB (RANK) expression in early, untreated RA synovium (D) decreased significantly following 8 weeks of MTX treatment (E) , as evaluated by semi-quantitative double-blind microscopic analysis (F) . Synovial osteoprotegerin (OPG) expression in early, untreated RA (G) is not changed following 8 weeks (H) of MTX treatment, as evaluated by semi-quantitative double-blind microscopic analysis (I) . *p
    Anti Human Rankl Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 97/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Abcam rabbit anti human rankl antibody
    Methotrexate decreases expression of <t>RANKL</t> and <t>RANK</t> in the rheumatoid synovium of early-untreated rheumatoid arthritis. Brown immunohistochemistry (diaminobenzidine) staining of cell surface markers in synovial biopsies of early rheumatoid arthritis (RA). Synovial receptor activator of the NF-κB ligand (RANKL) expression in early, untreated RA synovium (A) decreased significantly following 8 weeks of methotrexate (MTX) treatment (B) , as evaluated by semi-quantitative double-blind microscopic analysis (C) . Synovial receptor activator of NF-κB (RANK) expression in early, untreated RA synovium (D) decreased significantly following 8 weeks of MTX treatment (E) , as evaluated by semi-quantitative double-blind microscopic analysis (F) . Synovial osteoprotegerin (OPG) expression in early, untreated RA (G) is not changed following 8 weeks (H) of MTX treatment, as evaluated by semi-quantitative double-blind microscopic analysis (I) . *p
    Rabbit Anti Human Rankl Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 85/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems anti human rankl antibody
    Methotrexate decreases expression of <t>RANKL</t> and <t>RANK</t> in the rheumatoid synovium of early-untreated rheumatoid arthritis. Brown immunohistochemistry (diaminobenzidine) staining of cell surface markers in synovial biopsies of early rheumatoid arthritis (RA). Synovial receptor activator of the NF-κB ligand (RANKL) expression in early, untreated RA synovium (A) decreased significantly following 8 weeks of methotrexate (MTX) treatment (B) , as evaluated by semi-quantitative double-blind microscopic analysis (C) . Synovial receptor activator of NF-κB (RANK) expression in early, untreated RA synovium (D) decreased significantly following 8 weeks of MTX treatment (E) , as evaluated by semi-quantitative double-blind microscopic analysis (F) . Synovial osteoprotegerin (OPG) expression in early, untreated RA (G) is not changed following 8 weeks (H) of MTX treatment, as evaluated by semi-quantitative double-blind microscopic analysis (I) . *p
    Anti Human Rankl Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology anti human rankl ab
    Methotrexate decreases expression of <t>RANKL</t> and <t>RANK</t> in the rheumatoid synovium of early-untreated rheumatoid arthritis. Brown immunohistochemistry (diaminobenzidine) staining of cell surface markers in synovial biopsies of early rheumatoid arthritis (RA). Synovial receptor activator of the NF-κB ligand (RANKL) expression in early, untreated RA synovium (A) decreased significantly following 8 weeks of methotrexate (MTX) treatment (B) , as evaluated by semi-quantitative double-blind microscopic analysis (C) . Synovial receptor activator of NF-κB (RANK) expression in early, untreated RA synovium (D) decreased significantly following 8 weeks of MTX treatment (E) , as evaluated by semi-quantitative double-blind microscopic analysis (F) . Synovial osteoprotegerin (OPG) expression in early, untreated RA (G) is not changed following 8 weeks (H) of MTX treatment, as evaluated by semi-quantitative double-blind microscopic analysis (I) . *p
    Anti Human Rankl Ab, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam anti rankl mouse monoclonal antibody
    [ 99 mTc]Tc-Sestamibi uptake in breast cancer. ( A ) Image shows [ 99 mTc]Tc-Sestamibi uptake in a 55-year-old patient. ( B ) <t>RUNX2-</t> and <t>RANKL-positive</t> breast cancer cells. RANKL appears Texas-Red labeled, whereas RUNX2 is FITC-labeled. ( C ) Graph shows a significant association between lesion to non-lesion ratio and the number of breast osteoblast-like cells (BOLCs) (r 2 = 0.5056; p
    Anti Rankl Mouse Monoclonal Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Amgen recombinant human rankl
    [ 99 mTc]Tc-Sestamibi uptake in breast cancer. ( A ) Image shows [ 99 mTc]Tc-Sestamibi uptake in a 55-year-old patient. ( B ) <t>RUNX2-</t> and <t>RANKL-positive</t> breast cancer cells. RANKL appears Texas-Red labeled, whereas RUNX2 is FITC-labeled. ( C ) Graph shows a significant association between lesion to non-lesion ratio and the number of breast osteoblast-like cells (BOLCs) (r 2 = 0.5056; p
    Recombinant Human Rankl, supplied by Amgen, used in various techniques. Bioz Stars score: 89/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    FUJIFILM recombinant human rankl
    [ 99 mTc]Tc-Sestamibi uptake in breast cancer. ( A ) Image shows [ 99 mTc]Tc-Sestamibi uptake in a 55-year-old patient. ( B ) <t>RUNX2-</t> and <t>RANKL-positive</t> breast cancer cells. RANKL appears Texas-Red labeled, whereas RUNX2 is FITC-labeled. ( C ) Graph shows a significant association between lesion to non-lesion ratio and the number of breast osteoblast-like cells (BOLCs) (r 2 = 0.5056; p
    Recombinant Human Rankl, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rdi research diagnostics recombinant soluble human rankl
    CRAd armed with sOPG-Fc lyses cancer cells and inhibits osteoclast formation in vitro . MDA-MB-435 cells were infected with <t>Ad5-Δ24-sOPG-Fc-RGD,</t> Ad5-Δ24RGD or replication-defective vectors and grown for 9 days on inserts overlaying cocultures of murine osteoclast precursors and ST2 bone marrow stromal cells or human osteoclast precursors and <t>RANKL.</t> A , Cancer cells on the inserts were fixed and stained with crystal violet. The osteoclast marker protein TRAP5b was assayed by ELISA in the murine ( B ) or human ( C ) systems. Results are means ± SD of duplicate determinations. Representative results of 2 separate experiments are shown. Significant differences for sOPG-Fc-armed CRAd vs. uninfected (*) and unarmed CRAd (†) are indicated; P ≤ 0.05, determined by Student’s-Fisher t test. D, expression of sOPG-Fc in the medium of the murine system was quantified in an ELISA.
    Recombinant Soluble Human Rankl, supplied by rdi research diagnostics, used in various techniques. Bioz Stars score: 88/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc trance
    CRAd armed with sOPG-Fc lyses cancer cells and inhibits osteoclast formation in vitro . MDA-MB-435 cells were infected with <t>Ad5-Δ24-sOPG-Fc-RGD,</t> Ad5-Δ24RGD or replication-defective vectors and grown for 9 days on inserts overlaying cocultures of murine osteoclast precursors and ST2 bone marrow stromal cells or human osteoclast precursors and <t>RANKL.</t> A , Cancer cells on the inserts were fixed and stained with crystal violet. The osteoclast marker protein TRAP5b was assayed by ELISA in the murine ( B ) or human ( C ) systems. Results are means ± SD of duplicate determinations. Representative results of 2 separate experiments are shown. Significant differences for sOPG-Fc-armed CRAd vs. uninfected (*) and unarmed CRAd (†) are indicated; P ≤ 0.05, determined by Student’s-Fisher t test. D, expression of sOPG-Fc in the medium of the murine system was quantified in an ELISA.
    Trance, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Prospec human recombinant rankl
    CRAd armed with sOPG-Fc lyses cancer cells and inhibits osteoclast formation in vitro . MDA-MB-435 cells were infected with <t>Ad5-Δ24-sOPG-Fc-RGD,</t> Ad5-Δ24RGD or replication-defective vectors and grown for 9 days on inserts overlaying cocultures of murine osteoclast precursors and ST2 bone marrow stromal cells or human osteoclast precursors and <t>RANKL.</t> A , Cancer cells on the inserts were fixed and stained with crystal violet. The osteoclast marker protein TRAP5b was assayed by ELISA in the murine ( B ) or human ( C ) systems. Results are means ± SD of duplicate determinations. Representative results of 2 separate experiments are shown. Significant differences for sOPG-Fc-armed CRAd vs. uninfected (*) and unarmed CRAd (†) are indicated; P ≤ 0.05, determined by Student’s-Fisher t test. D, expression of sOPG-Fc in the medium of the murine system was quantified in an ELISA.
    Human Recombinant Rankl, supplied by Prospec, used in various techniques. Bioz Stars score: 93/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems human trance tnfsf11 rank l apc conjugated antibody
    CRAd armed with sOPG-Fc lyses cancer cells and inhibits osteoclast formation in vitro . MDA-MB-435 cells were infected with <t>Ad5-Δ24-sOPG-Fc-RGD,</t> Ad5-Δ24RGD or replication-defective vectors and grown for 9 days on inserts overlaying cocultures of murine osteoclast precursors and ST2 bone marrow stromal cells or human osteoclast precursors and <t>RANKL.</t> A , Cancer cells on the inserts were fixed and stained with crystal violet. The osteoclast marker protein TRAP5b was assayed by ELISA in the murine ( B ) or human ( C ) systems. Results are means ± SD of duplicate determinations. Representative results of 2 separate experiments are shown. Significant differences for sOPG-Fc-armed CRAd vs. uninfected (*) and unarmed CRAd (†) are indicated; P ≤ 0.05, determined by Student’s-Fisher t test. D, expression of sOPG-Fc in the medium of the murine system was quantified in an ELISA.
    Human Trance Tnfsf11 Rank L Apc Conjugated Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alexis Inc human rankl monoclonal antibody
    CRAd armed with sOPG-Fc lyses cancer cells and inhibits osteoclast formation in vitro . MDA-MB-435 cells were infected with <t>Ad5-Δ24-sOPG-Fc-RGD,</t> Ad5-Δ24RGD or replication-defective vectors and grown for 9 days on inserts overlaying cocultures of murine osteoclast precursors and ST2 bone marrow stromal cells or human osteoclast precursors and <t>RANKL.</t> A , Cancer cells on the inserts were fixed and stained with crystal violet. The osteoclast marker protein TRAP5b was assayed by ELISA in the murine ( B ) or human ( C ) systems. Results are means ± SD of duplicate determinations. Representative results of 2 separate experiments are shown. Significant differences for sOPG-Fc-armed CRAd vs. uninfected (*) and unarmed CRAd (†) are indicated; P ≤ 0.05, determined by Student’s-Fisher t test. D, expression of sOPG-Fc in the medium of the murine system was quantified in an ELISA.
    Human Rankl Monoclonal Antibody, supplied by Alexis Inc, used in various techniques. Bioz Stars score: 92/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    A : Immunohistochemical analysis of RANKL in C4-2 tumored tibiae of control, prevention, and treatment groups. Paraffin-embedded C4-2 tumored tibiae of the SCID mice were sectioned and stained for RANKL (negative control staining is shown in inset box). The tumor cells stained positively for RANKL. B : Blood was collected to determine serum PSA levels by IMx Total PSA assay. Serum PSA levels were not significantly different between the C4-2 control with PBS injections (control), C4-2 with a concomitant subcutaneous injection of huRANKL MAb (5 mg/kg once a week) at implantation (prevention group), or C4-2 with a subcutaneous injection of huRANKL MAb (5 mg/kg biweekly, starting 3 weeks after implantation of C4-2 cells) (treatment group). Results are presented as mean ± SE.

    Journal: BMC Cancer

    Article Title: Host-derived RANKL is responsible for osteolysis in a C4-2 human prostate cancer xenograft model of experimental bone metastases

    doi: 10.1186/1471-2407-7-148

    Figure Lengend Snippet: A : Immunohistochemical analysis of RANKL in C4-2 tumored tibiae of control, prevention, and treatment groups. Paraffin-embedded C4-2 tumored tibiae of the SCID mice were sectioned and stained for RANKL (negative control staining is shown in inset box). The tumor cells stained positively for RANKL. B : Blood was collected to determine serum PSA levels by IMx Total PSA assay. Serum PSA levels were not significantly different between the C4-2 control with PBS injections (control), C4-2 with a concomitant subcutaneous injection of huRANKL MAb (5 mg/kg once a week) at implantation (prevention group), or C4-2 with a subcutaneous injection of huRANKL MAb (5 mg/kg biweekly, starting 3 weeks after implantation of C4-2 cells) (treatment group). Results are presented as mean ± SE.

    Article Snippet: In the C4-2 tibial injection model of experimental bone metastases, the tumor cells express human RANKL, while the osteoblasts express murine RANKL.

    Techniques: Immunohistochemistry, Mouse Assay, Staining, Negative Control, Injection

    S100A12 stimulates the expression of mature-osteoclast markers. Human CD14 + monocytes were cultured in the presence of M-CSF (50 ng/ml), alone or in combination with RANKL (100 ng/ml) and with or without S100A12 (1,000 ng/ml) for 7–9 days. Levels of mRNA expression for nuclear factor of activated T cells c1 (NF-ATc1), TRAP (ACP5), calcitonin receptor (CALCR), integrin β3 (ITGβ3), cathepsin K (CTSK), and carbonic anhydrase II (CA2) were determined by qRT-PCR. Messenger RNA levels were normalized to GAPDH. Values are the mean and SD of 7 independent experiments. * = p

    Journal: PLoS ONE

    Article Title: S100A12 facilitates osteoclast differentiation from human monocytes

    doi: 10.1371/journal.pone.0204140

    Figure Lengend Snippet: S100A12 stimulates the expression of mature-osteoclast markers. Human CD14 + monocytes were cultured in the presence of M-CSF (50 ng/ml), alone or in combination with RANKL (100 ng/ml) and with or without S100A12 (1,000 ng/ml) for 7–9 days. Levels of mRNA expression for nuclear factor of activated T cells c1 (NF-ATc1), TRAP (ACP5), calcitonin receptor (CALCR), integrin β3 (ITGβ3), cathepsin K (CTSK), and carbonic anhydrase II (CA2) were determined by qRT-PCR. Messenger RNA levels were normalized to GAPDH. Values are the mean and SD of 7 independent experiments. * = p

    Article Snippet: Recombinant human M-CSF and recombinant human RANKL were purchased from R & D Systems (Minneapolis, MN).

    Techniques: Expressing, Cell Culture, Quantitative RT-PCR

    S100A12 has a similar stimulatory effect on human osteoclast differentiation as S100A8, and a weaker effect than S100A9. Human blood CD14 + monocytes were cultured with M-CSF (50 ng/ml), alone or in combination with RANKL (100 ng/ml) and S100A8 (1,000 ng/ml), S100A9 (1,000 ng/ml), or S100A12 (1,000 ng/ml). After 6 days of culture, the cells were fixed and stained for TRAP. (A) Representative images of TRAP staining (original magnification × 40). (B) TRAP-positive MNCs were counted. Values are the mean and SD of the results of 3 independent experiments. * = p

    Journal: PLoS ONE

    Article Title: S100A12 facilitates osteoclast differentiation from human monocytes

    doi: 10.1371/journal.pone.0204140

    Figure Lengend Snippet: S100A12 has a similar stimulatory effect on human osteoclast differentiation as S100A8, and a weaker effect than S100A9. Human blood CD14 + monocytes were cultured with M-CSF (50 ng/ml), alone or in combination with RANKL (100 ng/ml) and S100A8 (1,000 ng/ml), S100A9 (1,000 ng/ml), or S100A12 (1,000 ng/ml). After 6 days of culture, the cells were fixed and stained for TRAP. (A) Representative images of TRAP staining (original magnification × 40). (B) TRAP-positive MNCs were counted. Values are the mean and SD of the results of 3 independent experiments. * = p

    Article Snippet: Recombinant human M-CSF and recombinant human RANKL were purchased from R & D Systems (Minneapolis, MN).

    Techniques: Cell Culture, Staining

    S100A12 stimulates human osteoclast differentiation in a dose-dependent manner. Human blood CD14 + monocytes were cultured with M-CSF (50 ng/ml), alone or in combination with RANKL (100 ng/ml) and the indicated concentration of S100A12. Cells were fixed and stained for TRAP after 6 days of culture. (A) TRAP-positive multinucleated cells (MNCs) were counted. Values are the mean and SD of the results from 6 independent experiments. * = p

    Journal: PLoS ONE

    Article Title: S100A12 facilitates osteoclast differentiation from human monocytes

    doi: 10.1371/journal.pone.0204140

    Figure Lengend Snippet: S100A12 stimulates human osteoclast differentiation in a dose-dependent manner. Human blood CD14 + monocytes were cultured with M-CSF (50 ng/ml), alone or in combination with RANKL (100 ng/ml) and the indicated concentration of S100A12. Cells were fixed and stained for TRAP after 6 days of culture. (A) TRAP-positive multinucleated cells (MNCs) were counted. Values are the mean and SD of the results from 6 independent experiments. * = p

    Article Snippet: Recombinant human M-CSF and recombinant human RANKL were purchased from R & D Systems (Minneapolis, MN).

    Techniques: Cell Culture, Concentration Assay, Staining

    The effect of Hmox1 knockout in osteoclasts precursors on OCLs-specific genes expression. Bone marrow was isolated from HO-1 −/− and HO-1 +/+ mice. ( A , B ) BMCs-derived BMMs were stimulated with RANKL (50 ng/ml or 100 ng/ml) in the presence of 100 ng/ml M-CSF for 3 days. Alternatively, ( C , D ) BMCs-derived BMMs and ( E , F ) nBMCs-derived BMMs were replated and cultured in the presence of 50 ng/ml RANKL and 30 ng/ml M-CSF for 3 days. ( A , C , E ) NFAT-c1 and ( B , D , F ) cathepsin K relative expression (vs. EF-2). Quantitative PCR (n A,B = 9–10, n C,D = 7, n E,F = 17–20). Each bar represents the mean ± SEM. *p

    Journal: Scientific Reports

    Article Title: Various roles of heme oxygenase-1 in response of bone marrow macrophages to RANKL and in the early stage of osteoclastogenesis

    doi: 10.1038/s41598-018-29122-1

    Figure Lengend Snippet: The effect of Hmox1 knockout in osteoclasts precursors on OCLs-specific genes expression. Bone marrow was isolated from HO-1 −/− and HO-1 +/+ mice. ( A , B ) BMCs-derived BMMs were stimulated with RANKL (50 ng/ml or 100 ng/ml) in the presence of 100 ng/ml M-CSF for 3 days. Alternatively, ( C , D ) BMCs-derived BMMs and ( E , F ) nBMCs-derived BMMs were replated and cultured in the presence of 50 ng/ml RANKL and 30 ng/ml M-CSF for 3 days. ( A , C , E ) NFAT-c1 and ( B , D , F ) cathepsin K relative expression (vs. EF-2). Quantitative PCR (n A,B = 9–10, n C,D = 7, n E,F = 17–20). Each bar represents the mean ± SEM. *p

    Article Snippet: Reagents Recombinant human M-CSF and recombinant human RANKL were obtained from Sigma-Aldrich and were dissolved in water containing 0.1% BSA to a concentration of 10 μg/ml.

    Techniques: Knock-Out, Expressing, Isolation, Mouse Assay, Derivative Assay, Cell Culture, Real-time Polymerase Chain Reaction

    The effect of Nrf2 deficiency on OCLs markers expression. Bone marrow was isolated from Nrf2 −/− and Nrf2 +/+ mice. ( A – E ) Total BMCs-derived BMMs were stimulated with RANKL (50 or 100 ng/ml where indicated), and 2.5 μM sulforaphane where indicated, for 3 days in the presence of 100 ng/ml M-CSF. Alternatively, ( F ) total BMCs-derived BMMs or ( G ) nBMCs-derived BMMs were replated and treated with RANKL (50 ng/ml) in the presence of M-CSF (30 ng/ml). ( A , B ) Quantitative analysis and representative pictures (magnification 200x -upper panel and 400x -lower panel, scale bar – 50 µm) of TRAP + cells. TRAP staining (n = 6). ( C ) NFAT-c1, ( D ) integrin β3 and ( E ) cathepsin K relative expression (vs. EF-2). Quantitative PCR (n = 4–6). ( F , G ) Quantitative analysis of TRAP + cells. TRAP staining (n = 3–4). Each bar represents the mean ± SEM. *p

    Journal: Scientific Reports

    Article Title: Various roles of heme oxygenase-1 in response of bone marrow macrophages to RANKL and in the early stage of osteoclastogenesis

    doi: 10.1038/s41598-018-29122-1

    Figure Lengend Snippet: The effect of Nrf2 deficiency on OCLs markers expression. Bone marrow was isolated from Nrf2 −/− and Nrf2 +/+ mice. ( A – E ) Total BMCs-derived BMMs were stimulated with RANKL (50 or 100 ng/ml where indicated), and 2.5 μM sulforaphane where indicated, for 3 days in the presence of 100 ng/ml M-CSF. Alternatively, ( F ) total BMCs-derived BMMs or ( G ) nBMCs-derived BMMs were replated and treated with RANKL (50 ng/ml) in the presence of M-CSF (30 ng/ml). ( A , B ) Quantitative analysis and representative pictures (magnification 200x -upper panel and 400x -lower panel, scale bar – 50 µm) of TRAP + cells. TRAP staining (n = 6). ( C ) NFAT-c1, ( D ) integrin β3 and ( E ) cathepsin K relative expression (vs. EF-2). Quantitative PCR (n = 4–6). ( F , G ) Quantitative analysis of TRAP + cells. TRAP staining (n = 3–4). Each bar represents the mean ± SEM. *p

    Article Snippet: Reagents Recombinant human M-CSF and recombinant human RANKL were obtained from Sigma-Aldrich and were dissolved in water containing 0.1% BSA to a concentration of 10 μg/ml.

    Techniques: Expressing, Isolation, Mouse Assay, Derivative Assay, Staining, Real-time Polymerase Chain Reaction

    CoPPIX and hemin effect on osteoclasts-specific markers. ( A – D ) RAW264.7 were cultured with 50 ng/ml RANKL and 25 μM CoPPIX/hemin/SnPPIX or NaOH as a vehicle for ( A , C ) 9 h or ( B , D ) 48 h. ( A , B ) HO-1 and ( C , D ) NFAT-c1 relative expression (vs. EF-2). Quantitative PCR (n = 5–6). ( E – H ) Bone marrow was isolated from HO-1 −/− and HO-1 +/+ mice. ( E , F , G ) nBMCs-derived BMMs were replated and cultured for 3 days with 50 ng/ml RANKL and 30 ng/ml M-CSF in the presence of 5, 15 and 25 μM CoPPIX/hemin or DMSO as a vehicle or ( H ) total BMCs-derived BMMs were stimulated with 100 ng/ml RANKL and 100 ng/ml M-CSF for 3 days in the presence of 25 μM CoPPIX/hemin or DMSO as a vehicle. ( E ) HO-1 (in HO-1 +/+ cells), ( F) NFAT-C1 and ( G ) cathepsin K relative expression (vs. EF-2). Quantitative PCR (n E,F,G = 4). ( H ) Quantitative analysis of TRAP + cells ( > 2 nuclei). TRAP staining (n = 4). Each bar represents the mean ± SEM. *p

    Journal: Scientific Reports

    Article Title: Various roles of heme oxygenase-1 in response of bone marrow macrophages to RANKL and in the early stage of osteoclastogenesis

    doi: 10.1038/s41598-018-29122-1

    Figure Lengend Snippet: CoPPIX and hemin effect on osteoclasts-specific markers. ( A – D ) RAW264.7 were cultured with 50 ng/ml RANKL and 25 μM CoPPIX/hemin/SnPPIX or NaOH as a vehicle for ( A , C ) 9 h or ( B , D ) 48 h. ( A , B ) HO-1 and ( C , D ) NFAT-c1 relative expression (vs. EF-2). Quantitative PCR (n = 5–6). ( E – H ) Bone marrow was isolated from HO-1 −/− and HO-1 +/+ mice. ( E , F , G ) nBMCs-derived BMMs were replated and cultured for 3 days with 50 ng/ml RANKL and 30 ng/ml M-CSF in the presence of 5, 15 and 25 μM CoPPIX/hemin or DMSO as a vehicle or ( H ) total BMCs-derived BMMs were stimulated with 100 ng/ml RANKL and 100 ng/ml M-CSF for 3 days in the presence of 25 μM CoPPIX/hemin or DMSO as a vehicle. ( E ) HO-1 (in HO-1 +/+ cells), ( F) NFAT-C1 and ( G ) cathepsin K relative expression (vs. EF-2). Quantitative PCR (n E,F,G = 4). ( H ) Quantitative analysis of TRAP + cells ( > 2 nuclei). TRAP staining (n = 4). Each bar represents the mean ± SEM. *p

    Article Snippet: Reagents Recombinant human M-CSF and recombinant human RANKL were obtained from Sigma-Aldrich and were dissolved in water containing 0.1% BSA to a concentration of 10 μg/ml.

    Techniques: Cell Culture, Expressing, Real-time Polymerase Chain Reaction, Isolation, Mouse Assay, Derivative Assay, Staining

    The effect of Hmox1 knockout in osteoclasts precursors on TRAP+ cells formation. Bone marrow was isolated from HO-1 −/− and HO-1 +/+ mice. ( A , D ) BMCs-derived BMMs were stimulated with 100 ng/ml RANKL in the presence of 100 ng/ml M-CSF for 3 days. Alternatively, ( B , E ) BMCs-derived BMMs and ( C , F ) nBMCs-derived BMMs were replated and cultured in the presence of 50 ng/ml RANKL and 30 ng/ml M-CSF for 3 days. ( A , B , C ) Quantitative analysis and representative pictures (magnification 400x, scale bar – 50 µm) of TRAP + cells ( > 2 nuclei). TRAP staining (n A = 18–19, n B = 9–11, n C = 16–19). ( D , E , F ) TRAP concentration in the culture medium. TRAP ELISA (n = 5–6). Each bar represents the mean ± SEM. # p

    Journal: Scientific Reports

    Article Title: Various roles of heme oxygenase-1 in response of bone marrow macrophages to RANKL and in the early stage of osteoclastogenesis

    doi: 10.1038/s41598-018-29122-1

    Figure Lengend Snippet: The effect of Hmox1 knockout in osteoclasts precursors on TRAP+ cells formation. Bone marrow was isolated from HO-1 −/− and HO-1 +/+ mice. ( A , D ) BMCs-derived BMMs were stimulated with 100 ng/ml RANKL in the presence of 100 ng/ml M-CSF for 3 days. Alternatively, ( B , E ) BMCs-derived BMMs and ( C , F ) nBMCs-derived BMMs were replated and cultured in the presence of 50 ng/ml RANKL and 30 ng/ml M-CSF for 3 days. ( A , B , C ) Quantitative analysis and representative pictures (magnification 400x, scale bar – 50 µm) of TRAP + cells ( > 2 nuclei). TRAP staining (n A = 18–19, n B = 9–11, n C = 16–19). ( D , E , F ) TRAP concentration in the culture medium. TRAP ELISA (n = 5–6). Each bar represents the mean ± SEM. # p

    Article Snippet: Reagents Recombinant human M-CSF and recombinant human RANKL were obtained from Sigma-Aldrich and were dissolved in water containing 0.1% BSA to a concentration of 10 μg/ml.

    Techniques: Knock-Out, Isolation, Mouse Assay, Derivative Assay, Cell Culture, Staining, Concentration Assay, Enzyme-linked Immunosorbent Assay

    The effect of Hmox1 knockout in osteoclasts precursors on the formation of actin structures. Bone marrow was isolated from HO-1 −/− and HO-1 +/+ mice. nBMCs-derived BMMs were replated and cultured with 100 ng/ml RANKL for 5 days in the presence of 30 ng/ml M-CSF. Quantitative analysis (left) of multinucleated ( > 5 nuclei) cells with characterized actin structures and representative pictures (right, magnification 400x, scale bar –50 µm). Actin structures (green), nuclei (blue). White arrows indicate representative clusters of podosomes in the periphery of multinucleate cells. Immunofluorescence staining (n = 2–3, each from 12 fields of view). Each bar represents the mean ± SEM.

    Journal: Scientific Reports

    Article Title: Various roles of heme oxygenase-1 in response of bone marrow macrophages to RANKL and in the early stage of osteoclastogenesis

    doi: 10.1038/s41598-018-29122-1

    Figure Lengend Snippet: The effect of Hmox1 knockout in osteoclasts precursors on the formation of actin structures. Bone marrow was isolated from HO-1 −/− and HO-1 +/+ mice. nBMCs-derived BMMs were replated and cultured with 100 ng/ml RANKL for 5 days in the presence of 30 ng/ml M-CSF. Quantitative analysis (left) of multinucleated ( > 5 nuclei) cells with characterized actin structures and representative pictures (right, magnification 400x, scale bar –50 µm). Actin structures (green), nuclei (blue). White arrows indicate representative clusters of podosomes in the periphery of multinucleate cells. Immunofluorescence staining (n = 2–3, each from 12 fields of view). Each bar represents the mean ± SEM.

    Article Snippet: Reagents Recombinant human M-CSF and recombinant human RANKL were obtained from Sigma-Aldrich and were dissolved in water containing 0.1% BSA to a concentration of 10 μg/ml.

    Techniques: Knock-Out, Isolation, Mouse Assay, Derivative Assay, Cell Culture, Immunofluorescence, Staining

    The effect of HO-1 silencing in osteoclasts precursors on the expression of OCLs markers. Bone marrow was isolated from HO-1 +/+ mice. nBMCs-derived BMMs were replated and transfected with siRNA against HO-1 or scrambled control. One day after transfection fresh medium containing RANKL (50 ng/ml) and M-CSF (30 ng/ml) was added for the next 3 days. Quantitative PCR (n = 5). Each bar represents the mean ± SEM. # p

    Journal: Scientific Reports

    Article Title: Various roles of heme oxygenase-1 in response of bone marrow macrophages to RANKL and in the early stage of osteoclastogenesis

    doi: 10.1038/s41598-018-29122-1

    Figure Lengend Snippet: The effect of HO-1 silencing in osteoclasts precursors on the expression of OCLs markers. Bone marrow was isolated from HO-1 +/+ mice. nBMCs-derived BMMs were replated and transfected with siRNA against HO-1 or scrambled control. One day after transfection fresh medium containing RANKL (50 ng/ml) and M-CSF (30 ng/ml) was added for the next 3 days. Quantitative PCR (n = 5). Each bar represents the mean ± SEM. # p

    Article Snippet: Reagents Recombinant human M-CSF and recombinant human RANKL were obtained from Sigma-Aldrich and were dissolved in water containing 0.1% BSA to a concentration of 10 μg/ml.

    Techniques: Expressing, Isolation, Mouse Assay, Derivative Assay, Transfection, Real-time Polymerase Chain Reaction

    Microscopic image of a cultured osteoclast. Murine bone marrow macrophages were incubated for 82 h with RANKL (100 ng/mL) and M-CSF (10 ng/mL), resulting in osteoclast formation. After culturing, cells were fixed with 4% paraformaldehyde in phosphate buffer and stained with DAPI and Rhodamine B-conjugated phalloidin. Osteoclasts are multinucleated giant cells with a ring-shaped osteoclast-specific podosome belt termed the actin ring. Blue, nuclei; red, actin; scale bar = 50 µm.

    Journal: International Journal of Molecular Sciences

    Article Title: MicroRNAs: Potential Biomarkers and Therapeutic Targets for Alveolar Bone Loss in Periodontal Disease

    doi: 10.3390/ijms17081317

    Figure Lengend Snippet: Microscopic image of a cultured osteoclast. Murine bone marrow macrophages were incubated for 82 h with RANKL (100 ng/mL) and M-CSF (10 ng/mL), resulting in osteoclast formation. After culturing, cells were fixed with 4% paraformaldehyde in phosphate buffer and stained with DAPI and Rhodamine B-conjugated phalloidin. Osteoclasts are multinucleated giant cells with a ring-shaped osteoclast-specific podosome belt termed the actin ring. Blue, nuclei; red, actin; scale bar = 50 µm.

    Article Snippet: After three days, the medium was changed, and the cells were cultured in the presence of M-CSF (25 ng/mL) and recombinant human soluble RANKL (50 ng/mL) for an additional nine days.

    Techniques: Cell Culture, Incubation, Staining

    Expression levels of miR-155 and miR-223 during human osteoclast differentiation. Human peripheral blood CD14 + cells were treated with M-CSF (50 ng/mL) for three days, followed by RANKL (50 ng/mL) and M-CSF (25 ng/mL) for an additional nine days. Total RNAs were harvested at day 3 (macrophages) or day 12 (osteoclasts). The expression levels of miR-155 ( A ) and miR-223 ( B ) were then analyzed by quantitative RT-PCR. Values for each miRNA are expressed relative to those on day 3, which were set to 1. Quantification was performed using RNU6B as an endogenous control. The experiments were performed in triplicate. Data are presented as means ± SD. Student’s t -tests were performed to assess significant differences.

    Journal: International Journal of Molecular Sciences

    Article Title: MicroRNAs: Potential Biomarkers and Therapeutic Targets for Alveolar Bone Loss in Periodontal Disease

    doi: 10.3390/ijms17081317

    Figure Lengend Snippet: Expression levels of miR-155 and miR-223 during human osteoclast differentiation. Human peripheral blood CD14 + cells were treated with M-CSF (50 ng/mL) for three days, followed by RANKL (50 ng/mL) and M-CSF (25 ng/mL) for an additional nine days. Total RNAs were harvested at day 3 (macrophages) or day 12 (osteoclasts). The expression levels of miR-155 ( A ) and miR-223 ( B ) were then analyzed by quantitative RT-PCR. Values for each miRNA are expressed relative to those on day 3, which were set to 1. Quantification was performed using RNU6B as an endogenous control. The experiments were performed in triplicate. Data are presented as means ± SD. Student’s t -tests were performed to assess significant differences.

    Article Snippet: After three days, the medium was changed, and the cells were cultured in the presence of M-CSF (25 ng/mL) and recombinant human soluble RANKL (50 ng/mL) for an additional nine days.

    Techniques: Expressing, Quantitative RT-PCR

    A key osteoclastogenesis signaling cascade. Cited from [ 2 ]. The binding of M-CSF to its receptor, c-Fms, induces the transcription factor c-Fos, whereas the binding of RANKL to its receptor, RANK, leads to the recruitment of TNF-receptor-associated factor 6 (TRAF6), the main adapter molecule of RANK. TRAF6 activates nuclear factor κB (NF-κB) and mitogen-activated kinases including c-Jun N-terminal kinase (JNK). JNK activates the transcription factor c-Jun. RANKL/RANK also induces c-Fos to form AP-1, a heterodimeric transcription factor, with c-Jun. AP-1 and NF-κB then induce NFATc1, a master transcription factor that regulates osteoclast differentiation. NFATc1 works with other transcription factors, such as AP-1, PU.1, and microphthalmia-associated transcription factor (MITF) to induce various osteoclast-specific genes.

    Journal: International Journal of Molecular Sciences

    Article Title: MicroRNAs: Potential Biomarkers and Therapeutic Targets for Alveolar Bone Loss in Periodontal Disease

    doi: 10.3390/ijms17081317

    Figure Lengend Snippet: A key osteoclastogenesis signaling cascade. Cited from [ 2 ]. The binding of M-CSF to its receptor, c-Fms, induces the transcription factor c-Fos, whereas the binding of RANKL to its receptor, RANK, leads to the recruitment of TNF-receptor-associated factor 6 (TRAF6), the main adapter molecule of RANK. TRAF6 activates nuclear factor κB (NF-κB) and mitogen-activated kinases including c-Jun N-terminal kinase (JNK). JNK activates the transcription factor c-Jun. RANKL/RANK also induces c-Fos to form AP-1, a heterodimeric transcription factor, with c-Jun. AP-1 and NF-κB then induce NFATc1, a master transcription factor that regulates osteoclast differentiation. NFATc1 works with other transcription factors, such as AP-1, PU.1, and microphthalmia-associated transcription factor (MITF) to induce various osteoclast-specific genes.

    Article Snippet: After three days, the medium was changed, and the cells were cultured in the presence of M-CSF (25 ng/mL) and recombinant human soluble RANKL (50 ng/mL) for an additional nine days.

    Techniques: Binding Assay

    Inhibition of osteoclastogenesis by ASMSCs compared with HDMSCs. CD14 + monocytes were cultured with HDMSCs or ASMSCs in the presence of M-CSF and RANKL. a Representative images of TRAP staining of osteoclasts co-cultured at different time points (× 100). b The number of TRAP + osteoclasts in each well from cultures at different time points is shown. c Representative images of osteoclasts stained with FITC-phalloidin at different time points (× 200). d Representative images for bone resorption assays at different time points ( × 200). Cells cultured with HDMSCs or ASMSCs on bovine cortical slides were stained with toluidine blue. e Pit formation on each slide was assessed. f mRNA expression levels of TRAP, CTSK, and NFATc1 in osteoclasts were determined by qPCR on day 9. g Protein levels of TRAP, CTSK, and NFATc1 in osteoclasts were determined by western blot analyses on day 9. h Quantitative data of TRAP, CTSK, and NFATc1 protein levels determined by western blot analyses are shown. i Activation of signaling pathways involved in osteoclastogenesis was determined by western blot analyses on day 9. j Quantitative data for activation of signaling pathways determined by western blot analyses are shown. Values are the mean ± SD of 30 samples per group. The results represent three independent experiments. *, p

    Journal: Cell Death & Disease

    Article Title: Abnormal inhibition of osteoclastogenesis by mesenchymal stem cells through the miR-4284/CXCL5 axis in ankylosing spondylitis

    doi: 10.1038/s41419-019-1448-x

    Figure Lengend Snippet: Inhibition of osteoclastogenesis by ASMSCs compared with HDMSCs. CD14 + monocytes were cultured with HDMSCs or ASMSCs in the presence of M-CSF and RANKL. a Representative images of TRAP staining of osteoclasts co-cultured at different time points (× 100). b The number of TRAP + osteoclasts in each well from cultures at different time points is shown. c Representative images of osteoclasts stained with FITC-phalloidin at different time points (× 200). d Representative images for bone resorption assays at different time points ( × 200). Cells cultured with HDMSCs or ASMSCs on bovine cortical slides were stained with toluidine blue. e Pit formation on each slide was assessed. f mRNA expression levels of TRAP, CTSK, and NFATc1 in osteoclasts were determined by qPCR on day 9. g Protein levels of TRAP, CTSK, and NFATc1 in osteoclasts were determined by western blot analyses on day 9. h Quantitative data of TRAP, CTSK, and NFATc1 protein levels determined by western blot analyses are shown. i Activation of signaling pathways involved in osteoclastogenesis was determined by western blot analyses on day 9. j Quantitative data for activation of signaling pathways determined by western blot analyses are shown. Values are the mean ± SD of 30 samples per group. The results represent three independent experiments. *, p

    Article Snippet: For induction of osteoclast differentiation, CD14+ monocytes (2.5 × 105 /cm2 ) were cultured in α-minimum essential medium containing 10% fetal bovine serum supplemented with 25 ng/mL recombinant human M-CSF and 50 ng/mL recombinant human RANKL (both from Peprotech).

    Techniques: Inhibition, Cell Culture, Staining, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Activation Assay

    Effect of sh-CXCL5 on the inhibition of osteoclastogenesis by MSCs. CD14 + monocytes were cultured with HDMSCs or ASMSCs after knocking down CXCL5 in the presence of M-CSF and RANKL. a Secretion of CXCL5 from MSCs was detected by ELISA on day 3 after transfection with sh-CXCL5. b Representative images of TRAP staining (× 100). c The number of TRAP + osteoclasts in each well is shown. d Representative images of F-actin assays (× 200). e Representative images of bone resorption assays (× 200). f Pit formation on each slide was assessed. Data are presented as the mean ± SD ( n = 30). The results represent three independent experiments. *, p

    Journal: Cell Death & Disease

    Article Title: Abnormal inhibition of osteoclastogenesis by mesenchymal stem cells through the miR-4284/CXCL5 axis in ankylosing spondylitis

    doi: 10.1038/s41419-019-1448-x

    Figure Lengend Snippet: Effect of sh-CXCL5 on the inhibition of osteoclastogenesis by MSCs. CD14 + monocytes were cultured with HDMSCs or ASMSCs after knocking down CXCL5 in the presence of M-CSF and RANKL. a Secretion of CXCL5 from MSCs was detected by ELISA on day 3 after transfection with sh-CXCL5. b Representative images of TRAP staining (× 100). c The number of TRAP + osteoclasts in each well is shown. d Representative images of F-actin assays (× 200). e Representative images of bone resorption assays (× 200). f Pit formation on each slide was assessed. Data are presented as the mean ± SD ( n = 30). The results represent three independent experiments. *, p

    Article Snippet: For induction of osteoclast differentiation, CD14+ monocytes (2.5 × 105 /cm2 ) were cultured in α-minimum essential medium containing 10% fetal bovine serum supplemented with 25 ng/mL recombinant human M-CSF and 50 ng/mL recombinant human RANKL (both from Peprotech).

    Techniques: Inhibition, Cell Culture, Enzyme-linked Immunosorbent Assay, Transfection, Staining

    Mechanism of effect of IL-23 on human osteoclastogenesis. In a culture of human peripheral blood mononuclear cells, IL-23 directly induced osteoclastogenesis even in the absence of exogenous soluble receptor activator of NF-κB ligand (sRANKL). IL-17 induced by IL-23 from human activated T cells is the crucial cytokine for osteoclastogenesis through the mechanism of the RANK–RANKL system and TNF-α; IL-17 acts on monocytes, resulting in TNF-α production, inducing the differentiation of monocytes into osteoclasts by cooperating with RANKL, although RANKL alone expressed on monocytes does not induce osteoclastogenesis by binding to RANK. Furthermore, considering the inductive effect of IL-17 and the inhibitory effect of IFN-γ, it is speculated that the higher the ratio of IL-17 to IFN-γ, the greater the number of osteoclasts induced by IL-23. Ab, antibody; OPG, osteoprotegerin.

    Journal: Arthritis Research & Therapy

    Article Title: IL-23 induces human osteoclastogenesis via IL-17 in vitro, and anti-IL-23 antibody attenuates collagen-induced arthritis in rats

    doi: 10.1186/ar2297

    Figure Lengend Snippet: Mechanism of effect of IL-23 on human osteoclastogenesis. In a culture of human peripheral blood mononuclear cells, IL-23 directly induced osteoclastogenesis even in the absence of exogenous soluble receptor activator of NF-κB ligand (sRANKL). IL-17 induced by IL-23 from human activated T cells is the crucial cytokine for osteoclastogenesis through the mechanism of the RANK–RANKL system and TNF-α; IL-17 acts on monocytes, resulting in TNF-α production, inducing the differentiation of monocytes into osteoclasts by cooperating with RANKL, although RANKL alone expressed on monocytes does not induce osteoclastogenesis by binding to RANK. Furthermore, considering the inductive effect of IL-17 and the inhibitory effect of IFN-γ, it is speculated that the higher the ratio of IL-17 to IFN-γ, the greater the number of osteoclasts induced by IL-23. Ab, antibody; OPG, osteoprotegerin.

    Article Snippet: Recombinant human soluble RANKL (sRANKL) was obtained from PeproTech (London, UK).

    Techniques: Binding Assay

    Effects of NGF upon osteoclast differentiation in vitro . Human peripheral mononuclear cells were isolated and cultured in the presence of MCSF with (A) or without (B) RANKL, and increasing concentrations of NGF, for 14 days. TRAP +ve cells were counted using four random fields of vision on four separate slides for each condition. RANKL alone significantly increased TRAP +ve cells as compared to MCSF alone (A). For RANKL treated cells, addition of NGF significantly decreased osteoclast numbers at 50 and 100 ng/ml. In the absence of RANKL (B), addition of NGF caused a small non-dose related significant increase in TRAP +ve cell number. (C) Examples of TRAP staining in the presence and absence of RANKL. Scale bar is 100 μm. Data are mean ± SEM, statistical analysis: one way ANOVA, * P

    Journal: Osteoarthritis and Cartilage

    Article Title: The anti-NGF antibody muMab 911 both prevents and reverses pain behaviour and subchondral osteoclast numbers in a rat model of osteoarthritis pain

    doi: 10.1016/j.joca.2016.05.015

    Figure Lengend Snippet: Effects of NGF upon osteoclast differentiation in vitro . Human peripheral mononuclear cells were isolated and cultured in the presence of MCSF with (A) or without (B) RANKL, and increasing concentrations of NGF, for 14 days. TRAP +ve cells were counted using four random fields of vision on four separate slides for each condition. RANKL alone significantly increased TRAP +ve cells as compared to MCSF alone (A). For RANKL treated cells, addition of NGF significantly decreased osteoclast numbers at 50 and 100 ng/ml. In the absence of RANKL (B), addition of NGF caused a small non-dose related significant increase in TRAP +ve cell number. (C) Examples of TRAP staining in the presence and absence of RANKL. Scale bar is 100 μm. Data are mean ± SEM, statistical analysis: one way ANOVA, * P

    Article Snippet: In brief, peripheral blood from healthy human donors was collected and blood monocytes were isolated from buffy coats by gradient centrifugation, monocytes were seeded onto glass coverslips within a 24-well culture plates, and cultured in growth media supplemented with human macrophage colony stimulating factor (MCSF; R & D Systems) and with 30 ng ml−1 of human RANKL (Santa Cruz), unless otherwise stated.

    Techniques: In Vitro, Isolation, Cell Culture, Staining

    Methotrexate decreases expression of RANKL and RANK in the rheumatoid synovium of early-untreated rheumatoid arthritis. Brown immunohistochemistry (diaminobenzidine) staining of cell surface markers in synovial biopsies of early rheumatoid arthritis (RA). Synovial receptor activator of the NF-κB ligand (RANKL) expression in early, untreated RA synovium (A) decreased significantly following 8 weeks of methotrexate (MTX) treatment (B) , as evaluated by semi-quantitative double-blind microscopic analysis (C) . Synovial receptor activator of NF-κB (RANK) expression in early, untreated RA synovium (D) decreased significantly following 8 weeks of MTX treatment (E) , as evaluated by semi-quantitative double-blind microscopic analysis (F) . Synovial osteoprotegerin (OPG) expression in early, untreated RA (G) is not changed following 8 weeks (H) of MTX treatment, as evaluated by semi-quantitative double-blind microscopic analysis (I) . *p

    Journal: Arthritis Research & Therapy

    Article Title: Synovial membrane immunohistology in early-untreated rheumatoid arthritis reveals high expression of catabolic bone markers that is modulated by methotrexate

    doi: 10.1186/ar4398

    Figure Lengend Snippet: Methotrexate decreases expression of RANKL and RANK in the rheumatoid synovium of early-untreated rheumatoid arthritis. Brown immunohistochemistry (diaminobenzidine) staining of cell surface markers in synovial biopsies of early rheumatoid arthritis (RA). Synovial receptor activator of the NF-κB ligand (RANKL) expression in early, untreated RA synovium (A) decreased significantly following 8 weeks of methotrexate (MTX) treatment (B) , as evaluated by semi-quantitative double-blind microscopic analysis (C) . Synovial receptor activator of NF-κB (RANK) expression in early, untreated RA synovium (D) decreased significantly following 8 weeks of MTX treatment (E) , as evaluated by semi-quantitative double-blind microscopic analysis (F) . Synovial osteoprotegerin (OPG) expression in early, untreated RA (G) is not changed following 8 weeks (H) of MTX treatment, as evaluated by semi-quantitative double-blind microscopic analysis (I) . *p

    Article Snippet: Immunohistochemistry evaluation Presence of RANKL, RANK and OPG was detected using the following monoclonal antibodies: anti-human RANKL antibody at a final concentration of 5 μg/ml (12A668; Abcam, Cambridge, UK), anti-human RANK antibody at a final concentration of 5 μg/ml (9A725; Abcam) and anti-human OPG antibody (MAB805; R & D Systems, Minneapolis, MN, USA) at a concentration of 2.5 μg/ml, using a previously published protocol [ ].

    Techniques: Expressing, Immunohistochemistry, Staining

    Methotrexate decreases the synovial RANKL/OPG ratio in early-untreated rheumatoid arthritis. Synovial receptor activator of the NF-κB ligand (RANKL)/osteoprotegerin (OPG) ratio as evaluated by semi-quantitative double-blind microscopic analysis. * P

    Journal: Arthritis Research & Therapy

    Article Title: Synovial membrane immunohistology in early-untreated rheumatoid arthritis reveals high expression of catabolic bone markers that is modulated by methotrexate

    doi: 10.1186/ar4398

    Figure Lengend Snippet: Methotrexate decreases the synovial RANKL/OPG ratio in early-untreated rheumatoid arthritis. Synovial receptor activator of the NF-κB ligand (RANKL)/osteoprotegerin (OPG) ratio as evaluated by semi-quantitative double-blind microscopic analysis. * P

    Article Snippet: Immunohistochemistry evaluation Presence of RANKL, RANK and OPG was detected using the following monoclonal antibodies: anti-human RANKL antibody at a final concentration of 5 μg/ml (12A668; Abcam, Cambridge, UK), anti-human RANK antibody at a final concentration of 5 μg/ml (9A725; Abcam) and anti-human OPG antibody (MAB805; R & D Systems, Minneapolis, MN, USA) at a concentration of 2.5 μg/ml, using a previously published protocol [ ].

    Techniques:

    Methotrexate decreases mRNA and protein expression of RANKL and RANK in rheumatoid arthritis synovial-derived fibroblasts. A significant decrease in the receptor activator of the NF-κB ligand (RANKL) mRNA levels (A) with no changes in osteoprotegerin (OPG) mRNA levels (B) in the presence of methotrexate (MTX). Representative blots and graphs showing decrease of cellular RANKL protein expression (C and D) with no changes in the cellular OPG expression (C and E) in the presence of MTX. Decrease of soluble RANKL protein expression (F) with no changes in the soluble OPG expression (G) in the presence of MTX, where results are quantified by enzyme-linked immunosorbent assay. *p

    Journal: Arthritis Research & Therapy

    Article Title: Synovial membrane immunohistology in early-untreated rheumatoid arthritis reveals high expression of catabolic bone markers that is modulated by methotrexate

    doi: 10.1186/ar4398

    Figure Lengend Snippet: Methotrexate decreases mRNA and protein expression of RANKL and RANK in rheumatoid arthritis synovial-derived fibroblasts. A significant decrease in the receptor activator of the NF-κB ligand (RANKL) mRNA levels (A) with no changes in osteoprotegerin (OPG) mRNA levels (B) in the presence of methotrexate (MTX). Representative blots and graphs showing decrease of cellular RANKL protein expression (C and D) with no changes in the cellular OPG expression (C and E) in the presence of MTX. Decrease of soluble RANKL protein expression (F) with no changes in the soluble OPG expression (G) in the presence of MTX, where results are quantified by enzyme-linked immunosorbent assay. *p

    Article Snippet: Immunohistochemistry evaluation Presence of RANKL, RANK and OPG was detected using the following monoclonal antibodies: anti-human RANKL antibody at a final concentration of 5 μg/ml (12A668; Abcam, Cambridge, UK), anti-human RANK antibody at a final concentration of 5 μg/ml (9A725; Abcam) and anti-human OPG antibody (MAB805; R & D Systems, Minneapolis, MN, USA) at a concentration of 2.5 μg/ml, using a previously published protocol [ ].

    Techniques: Expressing, Derivative Assay, Enzyme-linked Immunosorbent Assay

    [ 99 mTc]Tc-Sestamibi uptake in breast cancer. ( A ) Image shows [ 99 mTc]Tc-Sestamibi uptake in a 55-year-old patient. ( B ) RUNX2- and RANKL-positive breast cancer cells. RANKL appears Texas-Red labeled, whereas RUNX2 is FITC-labeled. ( C ) Graph shows a significant association between lesion to non-lesion ratio and the number of breast osteoblast-like cells (BOLCs) (r 2 = 0.5056; p

    Journal: Journal of Clinical Medicine

    Article Title: Breast-Specific Gamma Imaging with [99mTc]Tc-Sestamibi: An In Vivo Analysis for Early Identification of Breast Cancer Lesions Expressing Bone Biomarkers

    doi: 10.3390/jcm9030747

    Figure Lengend Snippet: [ 99 mTc]Tc-Sestamibi uptake in breast cancer. ( A ) Image shows [ 99 mTc]Tc-Sestamibi uptake in a 55-year-old patient. ( B ) RUNX2- and RANKL-positive breast cancer cells. RANKL appears Texas-Red labeled, whereas RUNX2 is FITC-labeled. ( C ) Graph shows a significant association between lesion to non-lesion ratio and the number of breast osteoblast-like cells (BOLCs) (r 2 = 0.5056; p

    Article Snippet: Afterwards, sections were incubated with the primary antibodies diluted 1:100 for 60 min at room temperature: anti-RUNX2 mouse monoclonal antibody (clone 1D8, Novus Biologicals, Centennial, CO 80112, USA), anti-RANKL mouse monoclonal antibody (clone 12A668, AbCam, Cambridge, UK) anti-Ki67 rabbit monoclonal antibody (clone 30-9, Ventana, Tucson, USA), anti-ER rabbit monoclonal antibody (clone SP1, Novus Biologicals, Centennial, CO 80112, USA), anti-BMP-2 mouse monoclonal antibody (clone 9E10G12; Novus Biologicals, Centennial, CO 80112, USA), and anti-PTX3 rat monoclonal antibody (clone MNB1; Abcam, UK).

    Techniques: Labeling

    CRAd armed with sOPG-Fc lyses cancer cells and inhibits osteoclast formation in vitro . MDA-MB-435 cells were infected with Ad5-Δ24-sOPG-Fc-RGD, Ad5-Δ24RGD or replication-defective vectors and grown for 9 days on inserts overlaying cocultures of murine osteoclast precursors and ST2 bone marrow stromal cells or human osteoclast precursors and RANKL. A , Cancer cells on the inserts were fixed and stained with crystal violet. The osteoclast marker protein TRAP5b was assayed by ELISA in the murine ( B ) or human ( C ) systems. Results are means ± SD of duplicate determinations. Representative results of 2 separate experiments are shown. Significant differences for sOPG-Fc-armed CRAd vs. uninfected (*) and unarmed CRAd (†) are indicated; P ≤ 0.05, determined by Student’s-Fisher t test. D, expression of sOPG-Fc in the medium of the murine system was quantified in an ELISA.

    Journal: Cancer gene therapy

    Article Title: Arming a Replicating Adenovirus with Osteoprotegerin Reduces the Tumor Burden in a Murine Model of Osteolytic Bone Metastases of Breast Cancer

    doi: 10.1038/cgt.2010.47

    Figure Lengend Snippet: CRAd armed with sOPG-Fc lyses cancer cells and inhibits osteoclast formation in vitro . MDA-MB-435 cells were infected with Ad5-Δ24-sOPG-Fc-RGD, Ad5-Δ24RGD or replication-defective vectors and grown for 9 days on inserts overlaying cocultures of murine osteoclast precursors and ST2 bone marrow stromal cells or human osteoclast precursors and RANKL. A , Cancer cells on the inserts were fixed and stained with crystal violet. The osteoclast marker protein TRAP5b was assayed by ELISA in the murine ( B ) or human ( C ) systems. Results are means ± SD of duplicate determinations. Representative results of 2 separate experiments are shown. Significant differences for sOPG-Fc-armed CRAd vs. uninfected (*) and unarmed CRAd (†) are indicated; P ≤ 0.05, determined by Student’s-Fisher t test. D, expression of sOPG-Fc in the medium of the murine system was quantified in an ELISA.

    Article Snippet: Forty-eight hours postinfection of MDA-MB-231 cells with Ad5-Δ24-sOPG-Fc or Ad5-Δ24-sOPG-Fc-RGD, medium was harvested and incubated in the presence or absence of recombinant soluble human RANKL.

    Techniques: In Vitro, Multiple Displacement Amplification, Infection, Staining, Marker, Enzyme-linked Immunosorbent Assay, Expressing

    Characterization of armed CRAds. A, B, MDA-MB-231 cells were infected with Ad5-Δ24-sOPG-Fc, Ad5-Δ24-sOPG-Fc-RGD or Adwt300. At the indicated times, cellular RNA was subjected to QRT-PCR to detect expression of: A, the sOPG-Fc gene (cells infected with Ad5-Δ24-sOPG-Fc or Ad5-Δ24-sOPG-Fc-RGD) or the 14.7k gene (cells infected with Adwt300); and B, the ADP gene. The copy number was normalized to total cellular RNA. C, Secretion of sOPG-Fc. MDA-MB-231 cells were infected with Ad5-Δ24, Ad5-Δ24RGD, Ad5-Δ24-sOPG-Fc or Ad5-Δ24-sOPG-Fc-RGD. At the indicated times, medium was harvested, diluted 1:300 and expression of sOPG-Fc was detected in an ELISA. D , sOPG-Fc binds RANKL. Medium from MDA-MB-231 cells 48 h postinfection was incubated in the presence (+) or absence (−) of sRANKL and pulled down with protein G-agarose. Proteins were released and subjected to immunoblot using anti-RANKL primary antibody. C: sRANKL.

    Journal: Cancer gene therapy

    Article Title: Arming a Replicating Adenovirus with Osteoprotegerin Reduces the Tumor Burden in a Murine Model of Osteolytic Bone Metastases of Breast Cancer

    doi: 10.1038/cgt.2010.47

    Figure Lengend Snippet: Characterization of armed CRAds. A, B, MDA-MB-231 cells were infected with Ad5-Δ24-sOPG-Fc, Ad5-Δ24-sOPG-Fc-RGD or Adwt300. At the indicated times, cellular RNA was subjected to QRT-PCR to detect expression of: A, the sOPG-Fc gene (cells infected with Ad5-Δ24-sOPG-Fc or Ad5-Δ24-sOPG-Fc-RGD) or the 14.7k gene (cells infected with Adwt300); and B, the ADP gene. The copy number was normalized to total cellular RNA. C, Secretion of sOPG-Fc. MDA-MB-231 cells were infected with Ad5-Δ24, Ad5-Δ24RGD, Ad5-Δ24-sOPG-Fc or Ad5-Δ24-sOPG-Fc-RGD. At the indicated times, medium was harvested, diluted 1:300 and expression of sOPG-Fc was detected in an ELISA. D , sOPG-Fc binds RANKL. Medium from MDA-MB-231 cells 48 h postinfection was incubated in the presence (+) or absence (−) of sRANKL and pulled down with protein G-agarose. Proteins were released and subjected to immunoblot using anti-RANKL primary antibody. C: sRANKL.

    Article Snippet: Forty-eight hours postinfection of MDA-MB-231 cells with Ad5-Δ24-sOPG-Fc or Ad5-Δ24-sOPG-Fc-RGD, medium was harvested and incubated in the presence or absence of recombinant soluble human RANKL.

    Techniques: Multiple Displacement Amplification, Infection, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Incubation