collagenase type ii  (Millipore)

 
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    Name:
    Collagenase I
    Description:
    This product is provided as delivered and specified by the issuing Pharmacopoeia All information provided in support of this product including MSDS and any product information leaflets have been developed and issued under the Authority of the issuing Pharmacopoeia For further information and support please go to the website of the issuing Pharmacopoeia
    Catalog Number:
    1148089
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    Structured Review

    Millipore collagenase type ii
    Collagenase I
    This product is provided as delivered and specified by the issuing Pharmacopoeia All information provided in support of this product including MSDS and any product information leaflets have been developed and issued under the Authority of the issuing Pharmacopoeia For further information and support please go to the website of the issuing Pharmacopoeia
    https://www.bioz.com/result/collagenase type ii/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    collagenase type ii - by Bioz Stars, 2021-04
    97/100 stars

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    Incubation:

    Article Title: Accelerating skin wound healing by M-CSF through generating SSEA-1 and -3 stem cells in the injured sites
    Article Snippet: Isolation and culture of cells from mouse wounded skin or normal skin Four punch biopsies (6 mm) from wounded skin and eight punch biopsies from normal skin for each sacrificed mouse were taken. .. Tissue was minced into small pieces and incubated with type-1 collagenase (1 mg/ml in PBS, sigma) for 90 minutes in a shaker at 225 rpm and 37 °C. ..

    Article Title: CD105 (Endoglin)-Negative Murine Mesenchymal Stromal Cells Define a New Multipotent Subpopulation with Distinct Differentiation and Immunomodulatory Capacities
    Article Snippet: Cells were subsequently washed with PBS, fixed with 3.7% paraformaldehyde (15 minutes at room temperature) and stained with 1% cresyl violet. .. Flow cytometry Collagenase type I digests or cultured ASCs were incubated with 7AAD (Sigma-Aldrich) and 2.4G2 (eBioscience) and then stained with Abs specific for CD11b, CD29, CD31, CD34, CD44, CD45, CD49d, CD49f, CD61, CD73, CD105, CD140a, CD146, sca-1, MHC class I and MHC class II (all from eBioscience) or anti-TGF-βRI (ALK5) and anti-TGF-βRII Abs (R & D Systems, Minneapolis, MN). .. Cells were acquired and analyzed on a BD FACS Canto II using the FACS Diva software (BD Biosciences, Bedford, MA).

    Flow Cytometry:

    Article Title: Myeloid-Derived Lymphatic Endothelial Cell Progenitors Significantly Contribute to Lymphatic Metastasis in Clinical Breast Cancer
    Article Snippet: .. Flow Cytometry Analysis of CD11b+ Cells Isolated from Xenograft Tumors Orthotopic MDA-MB-231-Luc tumors harvested at a volume of 500 mm3 were digested by collagenase type III (225 U/mL) and hyaluronidase (100 U/mL), both from Sigma-Aldrich. .. Tumor-associated CD11b+ cells were isolated using anti-CD11b IgG-conjugated magnetic beads (Miltenyi Biotec).

    Article Title: CD105 (Endoglin)-Negative Murine Mesenchymal Stromal Cells Define a New Multipotent Subpopulation with Distinct Differentiation and Immunomodulatory Capacities
    Article Snippet: Cells were subsequently washed with PBS, fixed with 3.7% paraformaldehyde (15 minutes at room temperature) and stained with 1% cresyl violet. .. Flow cytometry Collagenase type I digests or cultured ASCs were incubated with 7AAD (Sigma-Aldrich) and 2.4G2 (eBioscience) and then stained with Abs specific for CD11b, CD29, CD31, CD34, CD44, CD45, CD49d, CD49f, CD61, CD73, CD105, CD140a, CD146, sca-1, MHC class I and MHC class II (all from eBioscience) or anti-TGF-βRI (ALK5) and anti-TGF-βRII Abs (R & D Systems, Minneapolis, MN). .. Cells were acquired and analyzed on a BD FACS Canto II using the FACS Diva software (BD Biosciences, Bedford, MA).

    Isolation:

    Article Title: Myeloid-Derived Lymphatic Endothelial Cell Progenitors Significantly Contribute to Lymphatic Metastasis in Clinical Breast Cancer
    Article Snippet: .. Flow Cytometry Analysis of CD11b+ Cells Isolated from Xenograft Tumors Orthotopic MDA-MB-231-Luc tumors harvested at a volume of 500 mm3 were digested by collagenase type III (225 U/mL) and hyaluronidase (100 U/mL), both from Sigma-Aldrich. .. Tumor-associated CD11b+ cells were isolated using anti-CD11b IgG-conjugated magnetic beads (Miltenyi Biotec).

    Article Title: Multilineage Differentiation Potential of Human Dental Pulp Stem Cells—Impact of 3D and Hypoxic Environment on Osteogenesis In Vitro
    Article Snippet: Subsequently, the pulp chamber was gently rinsed with phosphate-buffered saline (PBS; GE Healthcare Life Sciences HyClone Laboratories, Malborough, MA, USA) containing 100 IU/mL penicillin and 100 µg/mL streptomycin solutions (Gibco, ThermoFisher Scientific, Waltham, MA, USA) to wash out the remaining pulp tissue. .. Following the mechanical disruption, the isolated pulp tissue was subjected to further enzymatic digestion using a mixture of collagenase I (3 mg/mL, Sigma-Aldrich, St. Louis, MO, USA) and dispase (4 mg/mL, Sigma-Aldrich) for 30 min at 37 °C. .. The enzymes were inactivated by adding a complete cell culture medium (DMEM/F12 supplemented with 10% FBS, Sigma-Aldrich; and 100 IU/mL penicillin, 100 µg/mL streptomycin, Gibco, ThermoFisher Scientific).

    Multiple Displacement Amplification:

    Article Title: Myeloid-Derived Lymphatic Endothelial Cell Progenitors Significantly Contribute to Lymphatic Metastasis in Clinical Breast Cancer
    Article Snippet: .. Flow Cytometry Analysis of CD11b+ Cells Isolated from Xenograft Tumors Orthotopic MDA-MB-231-Luc tumors harvested at a volume of 500 mm3 were digested by collagenase type III (225 U/mL) and hyaluronidase (100 U/mL), both from Sigma-Aldrich. .. Tumor-associated CD11b+ cells were isolated using anti-CD11b IgG-conjugated magnetic beads (Miltenyi Biotec).

    Concentration Assay:

    Article Title: MMP-Sensitive PEG Diacrylate Hydrogels with Spatial Variations in Matrix Properties Stimulate Directional Vascular Sprout Formation
    Article Snippet: PBFP gradient hydrogels and their bulk equivalents were sectioned as outlined above and swollen in deionized water for 36 hours before being transferred to pre-weighed tubes and incubated in 10 mM HEPES buffered saline (HBS; 10 mM HEPES sodium salt and 137 mM NaCl) with 1 mM CaCl2 (pH 7.4) overnight. .. Collagenase-1A (Sigma Aldrich) at a concentration of 1 µg/mL was added to the hydrogel sections and the swollen weight of each section was recorded every two hours until complete hydrogel degradation was achieved. ..

    Mouse Assay:

    Article Title: Activation of dopamine D1 receptor decreased NLRP3-mediated inflammation in intracerebral hemorrhage mice
    Article Snippet: .. In this study, we used collagenase-induced ICH mice model to investigate whether A68930 (DRD1 specific agonist, Sigma) can improve neurological outcome through inhibition of NLRP3-mediated inflammation. .. Animals A total of 106 (excluding 3 mice that died because of anesthetic overdose) male CD1 mice (Charles River, Wilmington, MA, USA) weighing 30 to 34 g were used in these experiments.

    Inhibition:

    Article Title: Activation of dopamine D1 receptor decreased NLRP3-mediated inflammation in intracerebral hemorrhage mice
    Article Snippet: .. In this study, we used collagenase-induced ICH mice model to investigate whether A68930 (DRD1 specific agonist, Sigma) can improve neurological outcome through inhibition of NLRP3-mediated inflammation. .. Animals A total of 106 (excluding 3 mice that died because of anesthetic overdose) male CD1 mice (Charles River, Wilmington, MA, USA) weighing 30 to 34 g were used in these experiments.

    Cytometry:

    Article Title: CD105 (Endoglin)-Negative Murine Mesenchymal Stromal Cells Define a New Multipotent Subpopulation with Distinct Differentiation and Immunomodulatory Capacities
    Article Snippet: Cells were subsequently washed with PBS, fixed with 3.7% paraformaldehyde (15 minutes at room temperature) and stained with 1% cresyl violet. .. Flow cytometry Collagenase type I digests or cultured ASCs were incubated with 7AAD (Sigma-Aldrich) and 2.4G2 (eBioscience) and then stained with Abs specific for CD11b, CD29, CD31, CD34, CD44, CD45, CD49d, CD49f, CD61, CD73, CD105, CD140a, CD146, sca-1, MHC class I and MHC class II (all from eBioscience) or anti-TGF-βRI (ALK5) and anti-TGF-βRII Abs (R & D Systems, Minneapolis, MN). .. Cells were acquired and analyzed on a BD FACS Canto II using the FACS Diva software (BD Biosciences, Bedford, MA).

    Cell Culture:

    Article Title: CD105 (Endoglin)-Negative Murine Mesenchymal Stromal Cells Define a New Multipotent Subpopulation with Distinct Differentiation and Immunomodulatory Capacities
    Article Snippet: Cells were subsequently washed with PBS, fixed with 3.7% paraformaldehyde (15 minutes at room temperature) and stained with 1% cresyl violet. .. Flow cytometry Collagenase type I digests or cultured ASCs were incubated with 7AAD (Sigma-Aldrich) and 2.4G2 (eBioscience) and then stained with Abs specific for CD11b, CD29, CD31, CD34, CD44, CD45, CD49d, CD49f, CD61, CD73, CD105, CD140a, CD146, sca-1, MHC class I and MHC class II (all from eBioscience) or anti-TGF-βRI (ALK5) and anti-TGF-βRII Abs (R & D Systems, Minneapolis, MN). .. Cells were acquired and analyzed on a BD FACS Canto II using the FACS Diva software (BD Biosciences, Bedford, MA).

    Staining:

    Article Title: CD105 (Endoglin)-Negative Murine Mesenchymal Stromal Cells Define a New Multipotent Subpopulation with Distinct Differentiation and Immunomodulatory Capacities
    Article Snippet: Cells were subsequently washed with PBS, fixed with 3.7% paraformaldehyde (15 minutes at room temperature) and stained with 1% cresyl violet. .. Flow cytometry Collagenase type I digests or cultured ASCs were incubated with 7AAD (Sigma-Aldrich) and 2.4G2 (eBioscience) and then stained with Abs specific for CD11b, CD29, CD31, CD34, CD44, CD45, CD49d, CD49f, CD61, CD73, CD105, CD140a, CD146, sca-1, MHC class I and MHC class II (all from eBioscience) or anti-TGF-βRI (ALK5) and anti-TGF-βRII Abs (R & D Systems, Minneapolis, MN). .. Cells were acquired and analyzed on a BD FACS Canto II using the FACS Diva software (BD Biosciences, Bedford, MA).

    Injection:

    Article Title: Overexpression of mechanical sensitive miR-337-3p alleviates ectopic ossification in rat tendinopathy model via targeting IRS1 and Nox4 of tendon-derived stem cells
    Article Snippet: .. Bacterial collagenase I (20 μl of 0.015 mg/ml in saline; Sigma-Aldrich) or saline was injected into the patellar tendon intratendinously with a 29-gauge needle in one limb. .. Three days later, 50 μl rno-miR-337-3p-overexpressing lentivirus (1 × 109 TU/ml) or negative control lentivirus (GenePharma) was injected into the patellar tendon where collagenase I treated before.

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  • 93
    Millipore anti fgf 2 neutralizing antibody
    Effects of neutralizing endogenous fibroblast growth factor-2 <t>(FGF-2)</t> after subarachnoid hemorrhage (SAH). a Effects of anti-FGF-2 neutralizing antibody on neurobehavioral scores at 24 h after SAH (n = 6 per group). * P
    Anti Fgf 2 Neutralizing Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti fgf 2 neutralizing antibody/product/Millipore
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti fgf 2 neutralizing antibody - by Bioz Stars, 2021-04
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    99
    Millipore fgf 2
    PKCα mediates RANTES/CCL5-induced endothelial cell migration and vascular tube formation via syndecan-4. (A) Specificity of SDC-4 and pSDC-4 antibodies was checked using siRNA-negative control (SNC) or siRNA-SDC-4 (siRNA SDC-4) transfected cells by western blot analysis. HUV-EC-C transfected with GFP plasmid (control) or with SDC4WT-GFP (SDC4WT) were stimulated or not (U) by 3 nM RANTES/CCL5 (R) or 20 ng/ml <t>FGF-2</t> (F) and SDC-4 phosphorylation at Ser179 was evaluated by western blot. Upper panel, representative Western blot analysis. Lower panel, densitometry quantification of three independent experiments. pSDC-4 band intensity was normalized to SDC-4 one. Results of relative densitometry intensities (mean ± SEM) are expressed in arbitrary units (A.U.). * P
    Fgf 2, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fgf 2/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    fgf 2 - by Bioz Stars, 2021-04
    99/100 stars
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    Effects of neutralizing endogenous fibroblast growth factor-2 (FGF-2) after subarachnoid hemorrhage (SAH). a Effects of anti-FGF-2 neutralizing antibody on neurobehavioral scores at 24 h after SAH (n = 6 per group). * P

    Journal: Molecular neurobiology

    Article Title: FGF-2 attenuates neuronal apoptosis via FGFR3/PI3k/Akt signaling pathway after subarachnoid hemorrhage

    doi: 10.1007/s12035-019-01668-9

    Figure Lengend Snippet: Effects of neutralizing endogenous fibroblast growth factor-2 (FGF-2) after subarachnoid hemorrhage (SAH). a Effects of anti-FGF-2 neutralizing antibody on neurobehavioral scores at 24 h after SAH (n = 6 per group). * P

    Article Snippet: Anti-FGF-2 neutralizing antibody (05-117, EMD Millipore Inc., Temecula, CA) and negative control normal mouse IgG (12-371, EMD Millipore Inc., Temecula, CA) were administered i.c.v 1 h before SAH.

    Techniques:

    Effects of recombinant fibroblast growth factor-2 (rFGF-2) on long-term neuronal degeneration after subarachnoid hemorrhage (SAH). a  Representative Nissl staining pictures in different regions of hippocampus. Original magnification ×200.  b  Representative image demonstrating the location of interest area (small black boxes) in the left hippocampus.  c  Quantification of the Nissl-positive Neurons per area in dentate gyrus (DG), cornu ammonis (CA) 1, and CA3 at 28 days after modeling. Data are expressed as mean ± standard deviation (n = 8 per group). *  P

    Journal: Molecular neurobiology

    Article Title: FGF-2 attenuates neuronal apoptosis via FGFR3/PI3k/Akt signaling pathway after subarachnoid hemorrhage

    doi: 10.1007/s12035-019-01668-9

    Figure Lengend Snippet: Effects of recombinant fibroblast growth factor-2 (rFGF-2) on long-term neuronal degeneration after subarachnoid hemorrhage (SAH). a Representative Nissl staining pictures in different regions of hippocampus. Original magnification ×200. b Representative image demonstrating the location of interest area (small black boxes) in the left hippocampus. c Quantification of the Nissl-positive Neurons per area in dentate gyrus (DG), cornu ammonis (CA) 1, and CA3 at 28 days after modeling. Data are expressed as mean ± standard deviation (n = 8 per group). * P

    Article Snippet: Anti-FGF-2 neutralizing antibody (05-117, EMD Millipore Inc., Temecula, CA) and negative control normal mouse IgG (12-371, EMD Millipore Inc., Temecula, CA) were administered i.c.v 1 h before SAH.

    Techniques: Recombinant, Staining, Standard Deviation

    Expression of endogenous fibroblast growth factor (FGF)-2, FGF receptor (FGFR) 1, and FGFR3. a Representative western blot images and densitometric quantification of time-dependent expression of FGF-2, FGFR1, and FGFR3 after subarachnoid hemorrhage (SAH). Expression levels of each protein are expressed as a ratio of β-actin levels for normalization and as mean ± standard deviation. * P

    Journal: Molecular neurobiology

    Article Title: FGF-2 attenuates neuronal apoptosis via FGFR3/PI3k/Akt signaling pathway after subarachnoid hemorrhage

    doi: 10.1007/s12035-019-01668-9

    Figure Lengend Snippet: Expression of endogenous fibroblast growth factor (FGF)-2, FGF receptor (FGFR) 1, and FGFR3. a Representative western blot images and densitometric quantification of time-dependent expression of FGF-2, FGFR1, and FGFR3 after subarachnoid hemorrhage (SAH). Expression levels of each protein are expressed as a ratio of β-actin levels for normalization and as mean ± standard deviation. * P

    Article Snippet: Anti-FGF-2 neutralizing antibody (05-117, EMD Millipore Inc., Temecula, CA) and negative control normal mouse IgG (12-371, EMD Millipore Inc., Temecula, CA) were administered i.c.v 1 h before SAH.

    Techniques: Expressing, Western Blot, Standard Deviation

    Effects of recombinant fibroblast growth factor-2 (rFGF-2) on short-term neurobehavioral function and brain edema. a  9 μg rFGF-2 improved modified Garcia’s score and beam balance score at 24 h after subarachnoid hemorrhage (SAH). Data are expressed as median ± 25th-75th percentiles (n = 6-10 per group). †  P

    Journal: Molecular neurobiology

    Article Title: FGF-2 attenuates neuronal apoptosis via FGFR3/PI3k/Akt signaling pathway after subarachnoid hemorrhage

    doi: 10.1007/s12035-019-01668-9

    Figure Lengend Snippet: Effects of recombinant fibroblast growth factor-2 (rFGF-2) on short-term neurobehavioral function and brain edema. a 9 μg rFGF-2 improved modified Garcia’s score and beam balance score at 24 h after subarachnoid hemorrhage (SAH). Data are expressed as median ± 25th-75th percentiles (n = 6-10 per group). † P

    Article Snippet: Anti-FGF-2 neutralizing antibody (05-117, EMD Millipore Inc., Temecula, CA) and negative control normal mouse IgG (12-371, EMD Millipore Inc., Temecula, CA) were administered i.c.v 1 h before SAH.

    Techniques: Recombinant, Modification

    Effects of recombinant fibroblast growth factor-2 (rFGF-2) on short-term neuronal degeneration after subarachnoid hemorrhage (SAH). a  Representative images of Fluoro-Jade C (FJC)-positive cells in the left temporal cerebral cortex at 24 h after modeling. Scale bar = 50 μm.  b  Representative image demonstrating the location of staining (small black box).  c  Quantitative analysis of FJC-positive cells per mm 2  in the left cerebral cortex at 24 h after modeling. Data are expressed as mean ± standard deviation (n = 4 per group). *  P

    Journal: Molecular neurobiology

    Article Title: FGF-2 attenuates neuronal apoptosis via FGFR3/PI3k/Akt signaling pathway after subarachnoid hemorrhage

    doi: 10.1007/s12035-019-01668-9

    Figure Lengend Snippet: Effects of recombinant fibroblast growth factor-2 (rFGF-2) on short-term neuronal degeneration after subarachnoid hemorrhage (SAH). a Representative images of Fluoro-Jade C (FJC)-positive cells in the left temporal cerebral cortex at 24 h after modeling. Scale bar = 50 μm. b Representative image demonstrating the location of staining (small black box). c Quantitative analysis of FJC-positive cells per mm 2 in the left cerebral cortex at 24 h after modeling. Data are expressed as mean ± standard deviation (n = 4 per group). * P

    Article Snippet: Anti-FGF-2 neutralizing antibody (05-117, EMD Millipore Inc., Temecula, CA) and negative control normal mouse IgG (12-371, EMD Millipore Inc., Temecula, CA) were administered i.c.v 1 h before SAH.

    Techniques: Recombinant, Staining, Standard Deviation

    Effects of recombinant fibroblast growth factor-2 (rFGF-2) on short-term apoptotic marker after subarachnoid hemorrhage (SAH). a  Representative images demonstrating double immunofluorescent staining for cleaved caspase 3 and neuron-specific nuclear protein (NeuN) in the left temporal cerebral cortex at 24 h after modeling. Scale bar = 50 μm.  b  Representative image demonstrating the location of staining (small black box).  c  Quantitative analysis of number of cleaved caspase-3-positive cells per mm 2  in the left cerebral cortex at 24 h after modeling. Data are expressed as mean ± standard deviation (n = 4 per group). * P

    Journal: Molecular neurobiology

    Article Title: FGF-2 attenuates neuronal apoptosis via FGFR3/PI3k/Akt signaling pathway after subarachnoid hemorrhage

    doi: 10.1007/s12035-019-01668-9

    Figure Lengend Snippet: Effects of recombinant fibroblast growth factor-2 (rFGF-2) on short-term apoptotic marker after subarachnoid hemorrhage (SAH). a Representative images demonstrating double immunofluorescent staining for cleaved caspase 3 and neuron-specific nuclear protein (NeuN) in the left temporal cerebral cortex at 24 h after modeling. Scale bar = 50 μm. b Representative image demonstrating the location of staining (small black box). c Quantitative analysis of number of cleaved caspase-3-positive cells per mm 2 in the left cerebral cortex at 24 h after modeling. Data are expressed as mean ± standard deviation (n = 4 per group). * P

    Article Snippet: Anti-FGF-2 neutralizing antibody (05-117, EMD Millipore Inc., Temecula, CA) and negative control normal mouse IgG (12-371, EMD Millipore Inc., Temecula, CA) were administered i.c.v 1 h before SAH.

    Techniques: Recombinant, Marker, Staining, Standard Deviation

    Effects of recombinant fibroblast growth factor-2 (rFGF-2) on long-term neurobehavioral function after subarachnoid hemorrhage (SAH). a  Rotarod tests of 5 RPM and 10 RPM. * P

    Journal: Molecular neurobiology

    Article Title: FGF-2 attenuates neuronal apoptosis via FGFR3/PI3k/Akt signaling pathway after subarachnoid hemorrhage

    doi: 10.1007/s12035-019-01668-9

    Figure Lengend Snippet: Effects of recombinant fibroblast growth factor-2 (rFGF-2) on long-term neurobehavioral function after subarachnoid hemorrhage (SAH). a Rotarod tests of 5 RPM and 10 RPM. * P

    Article Snippet: Anti-FGF-2 neutralizing antibody (05-117, EMD Millipore Inc., Temecula, CA) and negative control normal mouse IgG (12-371, EMD Millipore Inc., Temecula, CA) were administered i.c.v 1 h before SAH.

    Techniques: Recombinant

    PKCα mediates RANTES/CCL5-induced endothelial cell migration and vascular tube formation via syndecan-4. (A) Specificity of SDC-4 and pSDC-4 antibodies was checked using siRNA-negative control (SNC) or siRNA-SDC-4 (siRNA SDC-4) transfected cells by western blot analysis. HUV-EC-C transfected with GFP plasmid (control) or with SDC4WT-GFP (SDC4WT) were stimulated or not (U) by 3 nM RANTES/CCL5 (R) or 20 ng/ml FGF-2 (F) and SDC-4 phosphorylation at Ser179 was evaluated by western blot. Upper panel, representative Western blot analysis. Lower panel, densitometry quantification of three independent experiments. pSDC-4 band intensity was normalized to SDC-4 one. Results of relative densitometry intensities (mean ± SEM) are expressed in arbitrary units (A.U.). * P

    Journal: Biology Open

    Article Title: RANTES/CCL5 mediated-biological effects depend on the syndecan-4/PKCα signaling pathway

    doi: 10.1242/bio.20148227

    Figure Lengend Snippet: PKCα mediates RANTES/CCL5-induced endothelial cell migration and vascular tube formation via syndecan-4. (A) Specificity of SDC-4 and pSDC-4 antibodies was checked using siRNA-negative control (SNC) or siRNA-SDC-4 (siRNA SDC-4) transfected cells by western blot analysis. HUV-EC-C transfected with GFP plasmid (control) or with SDC4WT-GFP (SDC4WT) were stimulated or not (U) by 3 nM RANTES/CCL5 (R) or 20 ng/ml FGF-2 (F) and SDC-4 phosphorylation at Ser179 was evaluated by western blot. Upper panel, representative Western blot analysis. Lower panel, densitometry quantification of three independent experiments. pSDC-4 band intensity was normalized to SDC-4 one. Results of relative densitometry intensities (mean ± SEM) are expressed in arbitrary units (A.U.). * P

    Article Snippet: Crystal Violet (0.1%), TPA (0.5 µM, 12-O-tetradecanoylphorbol-13-acetate), FGF-2 (20 ng/ml, Fibroblast Growth Factor-basic) and PKCα/β1 inhibitor Gö6976 (1 µM) were from Sigma–Aldrich (Saint-Quentin Fallavier, France).

    Techniques: Migration, Negative Control, Transfection, Western Blot, Plasmid Preparation

    BK/B2R transactivates FGFR-1 and mediates its internalization. ( A ) HUVEC were treated with BK (1 μM) for 8, 18, and 24 h, and FGF-2 expression was evaluated using western blot analysis. Results were normalized to actin. Quantification was expressed as an arbitrary density unit (ADU). The results presented are representative of three independent experiments ( n = 3) with similar results. ( B ) HUVEC were treated with fasitibant (fas, 1 µM, 30 min), then stimulated with BK (1 μM) for 24 h, and FGF-2 expression was evaluated using western blot analysis. Results were normalized to actin. ( C ) HUVEC were treated with BK (0.1–1000 nM, 10 min), FGFR-1 was immunoprecipitated (IP), and its activation was investigated by anti-pTYR antibody. Results were normalized to FGFR-1. ( D , E ) HUVEC were treated with fasitibant (fas, 1 µM, 30 min), then stimulated with 1 μM BK (10 min), FGFR-2 and FGFR-1 were immunoprecipitated (IP), and its activation was investigated by anti-pTYR antibody. Results were normalized to FGFR-2 and FGFR-1, respectively. *** p

    Journal: International Journal of Molecular Sciences

    Article Title: Bradykinin B2 Receptor Contributes to Inflammatory Responses in Human Endothelial Cells by the Transactivation of the Fibroblast Growth Factor Receptor FGFR-1

    doi: 10.3390/ijms19092638

    Figure Lengend Snippet: BK/B2R transactivates FGFR-1 and mediates its internalization. ( A ) HUVEC were treated with BK (1 μM) for 8, 18, and 24 h, and FGF-2 expression was evaluated using western blot analysis. Results were normalized to actin. Quantification was expressed as an arbitrary density unit (ADU). The results presented are representative of three independent experiments ( n = 3) with similar results. ( B ) HUVEC were treated with fasitibant (fas, 1 µM, 30 min), then stimulated with BK (1 μM) for 24 h, and FGF-2 expression was evaluated using western blot analysis. Results were normalized to actin. ( C ) HUVEC were treated with BK (0.1–1000 nM, 10 min), FGFR-1 was immunoprecipitated (IP), and its activation was investigated by anti-pTYR antibody. Results were normalized to FGFR-1. ( D , E ) HUVEC were treated with fasitibant (fas, 1 µM, 30 min), then stimulated with 1 μM BK (10 min), FGFR-2 and FGFR-1 were immunoprecipitated (IP), and its activation was investigated by anti-pTYR antibody. Results were normalized to FGFR-2 and FGFR-1, respectively. *** p

    Article Snippet: Cells (3 × 105 /6 cm plate) were treated with BK (1 μM) with or without the pre-treatment with fasitibant (1 µM), SU5402 (1 μM), PP1 (500 nM) or SU566 (Src inhibitors) (10 μM), or STAT3 inhibitor VII (10 µM) for 30 min. pFRSα (Tyr196, #3864 Cell Signaling), FRSα (#MAB4069 R & D system), pERK1/2 (#4370 Cell Signaling), ERK1/2 (#9102 Cell Signaling), pSTAT3 (#9134 Cell Signaling), STAT3 (#9139 Cell Signaling), pAKT (#9271 Cell Signaling), AKT (#2920 Cell Signaling), pTYR (#9411 Cell Signaling), p-SRC (#6943 Cell Signaling), SRC (#2108 Cell Signaling), FGFR-1 (#9740 Cell Signaling), FGFR-2 (#3116 Cell Signaling), and FGF-2 (#05–118 Merk Millipore) expression were evaluated using western blot analysis [ ].

    Techniques: Expressing, Western Blot, Immunoprecipitation, Activation Assay

    BK-mediated ERK1/2-STAT3/FGF-2 signaling activation requires FGFR-1. ( A ) ERK1/2 and ( B ) STAT3 phosphorylation were evaluated using western blot analysis in HUVEC treated with SU5402 (1 μM, 30 min), then stimulated with BK (1 μM) for 15 min. Results were normalized to ERK1/2 and STAT3, respectively. ( C ) FGFR-1 expression evaluated using western blot analysis in HUVEC transfected with two different shRNA for FGFR-1 knock-down (Sh#1 and Sh#2). EV = empty vector. ( D , E ) Western blot analysis for ERK1/2 and STAT3 phosphorylation in HUVEC transfected with Sh#1 and Sh#2 and stimulated with BK (1 μM) for 15 min. ( F ) FGF-2 expression was evaluated in HUVEC treated with STAT3 inhibitor (10 μM, 30 min) and then stimulated with BK (1 μM) for 24 h. Actin was used as a loading control. The results presented are representative of three independent experiments ( n = 3) with similar results. Quantification was expressed as an arbitrary density unit (ADU). ** p

    Journal: International Journal of Molecular Sciences

    Article Title: Bradykinin B2 Receptor Contributes to Inflammatory Responses in Human Endothelial Cells by the Transactivation of the Fibroblast Growth Factor Receptor FGFR-1

    doi: 10.3390/ijms19092638

    Figure Lengend Snippet: BK-mediated ERK1/2-STAT3/FGF-2 signaling activation requires FGFR-1. ( A ) ERK1/2 and ( B ) STAT3 phosphorylation were evaluated using western blot analysis in HUVEC treated with SU5402 (1 μM, 30 min), then stimulated with BK (1 μM) for 15 min. Results were normalized to ERK1/2 and STAT3, respectively. ( C ) FGFR-1 expression evaluated using western blot analysis in HUVEC transfected with two different shRNA for FGFR-1 knock-down (Sh#1 and Sh#2). EV = empty vector. ( D , E ) Western blot analysis for ERK1/2 and STAT3 phosphorylation in HUVEC transfected with Sh#1 and Sh#2 and stimulated with BK (1 μM) for 15 min. ( F ) FGF-2 expression was evaluated in HUVEC treated with STAT3 inhibitor (10 μM, 30 min) and then stimulated with BK (1 μM) for 24 h. Actin was used as a loading control. The results presented are representative of three independent experiments ( n = 3) with similar results. Quantification was expressed as an arbitrary density unit (ADU). ** p

    Article Snippet: Cells (3 × 105 /6 cm plate) were treated with BK (1 μM) with or without the pre-treatment with fasitibant (1 µM), SU5402 (1 μM), PP1 (500 nM) or SU566 (Src inhibitors) (10 μM), or STAT3 inhibitor VII (10 µM) for 30 min. pFRSα (Tyr196, #3864 Cell Signaling), FRSα (#MAB4069 R & D system), pERK1/2 (#4370 Cell Signaling), ERK1/2 (#9102 Cell Signaling), pSTAT3 (#9134 Cell Signaling), STAT3 (#9139 Cell Signaling), pAKT (#9271 Cell Signaling), AKT (#2920 Cell Signaling), pTYR (#9411 Cell Signaling), p-SRC (#6943 Cell Signaling), SRC (#2108 Cell Signaling), FGFR-1 (#9740 Cell Signaling), FGFR-2 (#3116 Cell Signaling), and FGF-2 (#05–118 Merk Millipore) expression were evaluated using western blot analysis [ ].

    Techniques: Activation Assay, Western Blot, Expressing, Transfection, shRNA, Plasmid Preparation

    BK phosphorylates FGFR-1 despite the absence of FGF-2. ( A , B ) HUVEC were treated with anti-FGF-2 neutralizing antibody (6 μg/mL), then stimulated with FGF-2 (20 ng/mL), as a positive control or BK (1 μM) for 10 min. FGFR-1 was immunoprecipitated (IP), and its activation was investigated by anti-pTYR antibody. Results were normalized to FGFR-1. *** p

    Journal: International Journal of Molecular Sciences

    Article Title: Bradykinin B2 Receptor Contributes to Inflammatory Responses in Human Endothelial Cells by the Transactivation of the Fibroblast Growth Factor Receptor FGFR-1

    doi: 10.3390/ijms19092638

    Figure Lengend Snippet: BK phosphorylates FGFR-1 despite the absence of FGF-2. ( A , B ) HUVEC were treated with anti-FGF-2 neutralizing antibody (6 μg/mL), then stimulated with FGF-2 (20 ng/mL), as a positive control or BK (1 μM) for 10 min. FGFR-1 was immunoprecipitated (IP), and its activation was investigated by anti-pTYR antibody. Results were normalized to FGFR-1. *** p

    Article Snippet: Cells (3 × 105 /6 cm plate) were treated with BK (1 μM) with or without the pre-treatment with fasitibant (1 µM), SU5402 (1 μM), PP1 (500 nM) or SU566 (Src inhibitors) (10 μM), or STAT3 inhibitor VII (10 µM) for 30 min. pFRSα (Tyr196, #3864 Cell Signaling), FRSα (#MAB4069 R & D system), pERK1/2 (#4370 Cell Signaling), ERK1/2 (#9102 Cell Signaling), pSTAT3 (#9134 Cell Signaling), STAT3 (#9139 Cell Signaling), pAKT (#9271 Cell Signaling), AKT (#2920 Cell Signaling), pTYR (#9411 Cell Signaling), p-SRC (#6943 Cell Signaling), SRC (#2108 Cell Signaling), FGFR-1 (#9740 Cell Signaling), FGFR-2 (#3116 Cell Signaling), and FGF-2 (#05–118 Merk Millipore) expression were evaluated using western blot analysis [ ].

    Techniques: Positive Control, Immunoprecipitation, Activation Assay

    Schematic model of BK/B2R-FGF-2/FGFR-1 interaction in endothelial cells. The figure depicts the interaction between BK and FGF-2 signaling in endothelial cells. Dotted arrows indicate complex signaling pathways involving several second messengers.

    Journal: International Journal of Molecular Sciences

    Article Title: Bradykinin B2 Receptor Contributes to Inflammatory Responses in Human Endothelial Cells by the Transactivation of the Fibroblast Growth Factor Receptor FGFR-1

    doi: 10.3390/ijms19092638

    Figure Lengend Snippet: Schematic model of BK/B2R-FGF-2/FGFR-1 interaction in endothelial cells. The figure depicts the interaction between BK and FGF-2 signaling in endothelial cells. Dotted arrows indicate complex signaling pathways involving several second messengers.

    Article Snippet: Cells (3 × 105 /6 cm plate) were treated with BK (1 μM) with or without the pre-treatment with fasitibant (1 µM), SU5402 (1 μM), PP1 (500 nM) or SU566 (Src inhibitors) (10 μM), or STAT3 inhibitor VII (10 µM) for 30 min. pFRSα (Tyr196, #3864 Cell Signaling), FRSα (#MAB4069 R & D system), pERK1/2 (#4370 Cell Signaling), ERK1/2 (#9102 Cell Signaling), pSTAT3 (#9134 Cell Signaling), STAT3 (#9139 Cell Signaling), pAKT (#9271 Cell Signaling), AKT (#2920 Cell Signaling), pTYR (#9411 Cell Signaling), p-SRC (#6943 Cell Signaling), SRC (#2108 Cell Signaling), FGFR-1 (#9740 Cell Signaling), FGFR-2 (#3116 Cell Signaling), and FGF-2 (#05–118 Merk Millipore) expression were evaluated using western blot analysis [ ].

    Techniques: