vascular endothelial growth factor  (Cell Signaling Technology Inc)

 
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    Name:
    Vascular Endothelial Growth Factor VEGF
    Description:
    The human VEGF165 coding cDNA was subcloned into an expression vector and expressed in yeast The recombinant human VEGF 165 homodimer was purified and stored in PBS buffer pH 7 4 containing 0 1 BSA
    Catalog Number:
    9943
    Price:
    None
    Category:
    Cytokines
    Source:
    Human Recombinant Protein
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    Structured Review

    Cell Signaling Technology Inc vascular endothelial growth factor
    The human VEGF165 coding cDNA was subcloned into an expression vector and expressed in yeast The recombinant human VEGF 165 homodimer was purified and stored in PBS buffer pH 7 4 containing 0 1 BSA
    https://www.bioz.com/result/vascular endothelial growth factor/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    vascular endothelial growth factor - by Bioz Stars, 2021-03
    97/100 stars

    Images

    Related Articles

    Western Blot:

    Article Title: Augmentation of neovascularization in murine hindlimb ischemia by combined therapy with simvastatin and bone marrow-derived mesenchymal stem cells transplantation
    Article Snippet: Cells in which the nucleus was stained brown were defined as TUNEL-positive and the percentage of apoptotic cells per total number of cells was determined by two independent blinded investigators. .. Western blot analysis for the expression of VEGF protein in vivo Lysates from hind limb muscle tissue homogenates harvested at day 21 post-surgery were used for Western blot analysis as described previously[ ].Protein was analyzed using 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to nitrocellulose membranes (Bio-Rad).Membranes were then incubated with primary antibodies including VEGF (1:1000, Cell Signaling) and β-actin (1:5000, Sigma) at 4°C overnight respectively.The membranes were then incubated with peroxidase labeled secondary antibody (1:1000, Santa Cruz, USA) at 37°C for 2 hours. .. Signals were detected by enhanced chemiluminescence (Amersham, USA).

    Expressing:

    Article Title: Augmentation of neovascularization in murine hindlimb ischemia by combined therapy with simvastatin and bone marrow-derived mesenchymal stem cells transplantation
    Article Snippet: Cells in which the nucleus was stained brown were defined as TUNEL-positive and the percentage of apoptotic cells per total number of cells was determined by two independent blinded investigators. .. Western blot analysis for the expression of VEGF protein in vivo Lysates from hind limb muscle tissue homogenates harvested at day 21 post-surgery were used for Western blot analysis as described previously[ ].Protein was analyzed using 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to nitrocellulose membranes (Bio-Rad).Membranes were then incubated with primary antibodies including VEGF (1:1000, Cell Signaling) and β-actin (1:5000, Sigma) at 4°C overnight respectively.The membranes were then incubated with peroxidase labeled secondary antibody (1:1000, Santa Cruz, USA) at 37°C for 2 hours. .. Signals were detected by enhanced chemiluminescence (Amersham, USA).

    In Vivo:

    Article Title: Augmentation of neovascularization in murine hindlimb ischemia by combined therapy with simvastatin and bone marrow-derived mesenchymal stem cells transplantation
    Article Snippet: Cells in which the nucleus was stained brown were defined as TUNEL-positive and the percentage of apoptotic cells per total number of cells was determined by two independent blinded investigators. .. Western blot analysis for the expression of VEGF protein in vivo Lysates from hind limb muscle tissue homogenates harvested at day 21 post-surgery were used for Western blot analysis as described previously[ ].Protein was analyzed using 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to nitrocellulose membranes (Bio-Rad).Membranes were then incubated with primary antibodies including VEGF (1:1000, Cell Signaling) and β-actin (1:5000, Sigma) at 4°C overnight respectively.The membranes were then incubated with peroxidase labeled secondary antibody (1:1000, Santa Cruz, USA) at 37°C for 2 hours. .. Signals were detected by enhanced chemiluminescence (Amersham, USA).

    Nucleic Acid Electrophoresis:

    Article Title: Augmentation of neovascularization in murine hindlimb ischemia by combined therapy with simvastatin and bone marrow-derived mesenchymal stem cells transplantation
    Article Snippet: Cells in which the nucleus was stained brown were defined as TUNEL-positive and the percentage of apoptotic cells per total number of cells was determined by two independent blinded investigators. .. Western blot analysis for the expression of VEGF protein in vivo Lysates from hind limb muscle tissue homogenates harvested at day 21 post-surgery were used for Western blot analysis as described previously[ ].Protein was analyzed using 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to nitrocellulose membranes (Bio-Rad).Membranes were then incubated with primary antibodies including VEGF (1:1000, Cell Signaling) and β-actin (1:5000, Sigma) at 4°C overnight respectively.The membranes were then incubated with peroxidase labeled secondary antibody (1:1000, Santa Cruz, USA) at 37°C for 2 hours. .. Signals were detected by enhanced chemiluminescence (Amersham, USA).

    SDS Page:

    Article Title: Augmentation of neovascularization in murine hindlimb ischemia by combined therapy with simvastatin and bone marrow-derived mesenchymal stem cells transplantation
    Article Snippet: Cells in which the nucleus was stained brown were defined as TUNEL-positive and the percentage of apoptotic cells per total number of cells was determined by two independent blinded investigators. .. Western blot analysis for the expression of VEGF protein in vivo Lysates from hind limb muscle tissue homogenates harvested at day 21 post-surgery were used for Western blot analysis as described previously[ ].Protein was analyzed using 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to nitrocellulose membranes (Bio-Rad).Membranes were then incubated with primary antibodies including VEGF (1:1000, Cell Signaling) and β-actin (1:5000, Sigma) at 4°C overnight respectively.The membranes were then incubated with peroxidase labeled secondary antibody (1:1000, Santa Cruz, USA) at 37°C for 2 hours. .. Signals were detected by enhanced chemiluminescence (Amersham, USA).

    Incubation:

    Article Title: Augmentation of neovascularization in murine hindlimb ischemia by combined therapy with simvastatin and bone marrow-derived mesenchymal stem cells transplantation
    Article Snippet: Cells in which the nucleus was stained brown were defined as TUNEL-positive and the percentage of apoptotic cells per total number of cells was determined by two independent blinded investigators. .. Western blot analysis for the expression of VEGF protein in vivo Lysates from hind limb muscle tissue homogenates harvested at day 21 post-surgery were used for Western blot analysis as described previously[ ].Protein was analyzed using 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to nitrocellulose membranes (Bio-Rad).Membranes were then incubated with primary antibodies including VEGF (1:1000, Cell Signaling) and β-actin (1:5000, Sigma) at 4°C overnight respectively.The membranes were then incubated with peroxidase labeled secondary antibody (1:1000, Santa Cruz, USA) at 37°C for 2 hours. .. Signals were detected by enhanced chemiluminescence (Amersham, USA).

    Article Title: Increasing the expression of microRNA-126-5p in the temporal muscle can promote angiogenesis in the chronically ischemic brains of rats subjected to two-vessel occlusion plus encephalo-myo-synangiosis
    Article Snippet: Briefly, sections were fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS) for 5 min at room temperature, permeabilized, and blocked for 30 min with 0.1% Triton X-100 and 1% bovine serum albumin. .. Subsequently, the fixed sections were washed and incubated for 1 h with primary antibodies against vWF (1:200; Cell Signaling Technology, Shanghai, China), VEGF (1:200; Cell Signaling Technology), CD31 (1:100; Santa Cruz Biotechnology, Dallas, TX, USA), or eNOS (1:200; Cell Signaling Technology) overnight at 4°C. .. The sections were then washed three times with PBS and incubated with fluorescence-conjugated species-specific secondary antibodies.

    Article Title: LncRNA SNHG14 promotes hepatocellular carcinoma progression via H3K27 acetylation activated PABPC1 by PTEN signaling
    Article Snippet: For IHC analysis, paraffin sections were deparaffinized, rehydrated, and subjected to antigen retrieval. .. Slides were then blocked with 10% normal goat serum and incubated with anti-Ki67 (1:500, Abcam, Cambridge, MA, USA), anti-CD31 antibody (1:100, Thermo Fisher Scientific), PTEN (1:100, Cell Signaling Technology, Beverly, MA, USA), or VEGF (1:100, Cell Signaling Technology) at 4 °C overnight, followed by an incubation with biotinylated secondary antibody and streptavidin-HRP. .. Immunoreactivity was visualized using AEC solution (Thermo Fisher Scientific).

    Article Title: Effect of cycloxygenase-2 silencing on the malignant biological behavior of MCF-7 breast cancer cells
    Article Snippet: Protein from 10% SDS-PAGE gel was transferred to a polyvinylidene difluoride membrane following electrophoresis. .. Next, the primary rabbit monoclonal anti-COX-2 (1:500), anti-vascular endothelial growth factor (VEGF)-A (1:800), anti-VEGF-C (1:800) and anti-GAPDH (1:4,000) antibodies (Cell Signaling Technology, Inc., Danvers, MA, USA) were added and the mixture was incubated overnight at 4°C on a rocking platform. .. Subsequent to being washed, the membrane was added together with the horseradish peroxidase-conjugated secondary antibody (1:4,000) and incubated for 2 h. The membrane was then developed using an enhanced chemiluminescence system (Pierce Biotechnology, Inc., Rockford, IL, USA) and exposed to X-ray film.

    Labeling:

    Article Title: Augmentation of neovascularization in murine hindlimb ischemia by combined therapy with simvastatin and bone marrow-derived mesenchymal stem cells transplantation
    Article Snippet: Cells in which the nucleus was stained brown were defined as TUNEL-positive and the percentage of apoptotic cells per total number of cells was determined by two independent blinded investigators. .. Western blot analysis for the expression of VEGF protein in vivo Lysates from hind limb muscle tissue homogenates harvested at day 21 post-surgery were used for Western blot analysis as described previously[ ].Protein was analyzed using 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to nitrocellulose membranes (Bio-Rad).Membranes were then incubated with primary antibodies including VEGF (1:1000, Cell Signaling) and β-actin (1:5000, Sigma) at 4°C overnight respectively.The membranes were then incubated with peroxidase labeled secondary antibody (1:1000, Santa Cruz, USA) at 37°C for 2 hours. .. Signals were detected by enhanced chemiluminescence (Amersham, USA).

    Immunoprecipitation:

    Article Title: Paxillin regulates vascular endothelial growth factor A-induced in vitro angiogenesis of human umbilical vein endothelial cells
    Article Snippet: The coverslips were then washed with PBS, the cells were examined with a fluorescence microscope (Olympus, Tokyo, Japan) and images were captured with a DP70 digital camera (Olympus). .. Immunoprecipitation The HUVECs were grown to confluence and stimulated with 20 ng/ml VEGF-A (Cell Signaling Technology, Inc., Beverly, MA, USA) at 37°C for 0, 20, 40 and 60 min. .. The cells were then washed with ice-cold PBS and solubilized on ice with lysis buffer containing 150 mM NaCl, 10 mM Tris-HCl, (pH 7.5) and 1% Triton X-100 supplemented with a cocktail of phosphatase and proteinase inhibitors containing 1 mM vanadate, 10 mg/ml leupeptin, 10 mg/ml aprotinin, 1 mM phenylmethylsulfonyl fluoride and 0.36 mM phenanthroline.

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    Cell Signaling Technology Inc rabbit anti vegf
    Effect of curcumin and hypoxia on <t>IGF-1R,</t> p-Akt, p-Erk1/2, HIF-1α, <t>VEGF</t> protein expression in IGF-1R knockout HepG2 cells. (A) IGF-1R, p-Akt, p-Erk1/2, HIF-1α, VEGF protein expression were detected by western blot analysis. Graphic representation of relative density of (B) IGF-1R, (C) p-Akt, (D) p-Erk1/2, (E) HIF-1α and (F) VEGF protein levels, which were normalized to those of β-actin, Akt or Erk1/2, respectively. *P
    Rabbit Anti Vegf, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc vegfr 2 kinase activity
    Isomangiferin inhibits breast tumor growth and vascular endothelial growth factor receptor 2 <t>(VEGFR-2)</t> signaling pathway in vivo . Human breast cancer cells MDA-MB-231 were injected subcutanously into 5-week-old BALB/cA nude mice (2×10 6 per mouse). When subcutanous tumors grew to about 100 mm 3 , the mice were intraperitoneally treated with or without isomangiferin (10 mg/kg/day). (A) Photos for isomangiferin treated or non-treated tumors. (B) Isomangiferin supressed MDA-MB-231 xenografts growth. Tumor volume was recorded every 6 days and the tumor growth curve was drafted through Graphpad Prism 5 software package. Values are shown as mean±standard error of the mean (SEM) of three independent experiments. (C) Isomangiferin inhibited breast tumor growth as measured by tumor weight. Values are shown as mean±SEM of three independent experiments. (D) Immunohistochemical staining revealed that isomangiferin inhibited VEGFR-2 signaling pathway by blotting phosphorylated VEGFR-2 (p-VEGFR-2) and reducing CD31 expression. Tumor sections from isomangiferin-treated and isomangiferin-untreated groups were stained using p-VEGFR-2 and CD31 antibodies, and the number of positive cells was counted (IHC stain for p-VEGFR-2, CD31, ×400). * p
    Vegfr 2 Kinase Activity, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc vegf a
    Knockdown of paxillin inhibits the <t>VEGF-A-induced</t> migration of <t>HUVECs.</t> Cells were cultured on plates following either mock-transfection or transfection with scramble siRNA or siPXN-1, respectively. HUVECs (2×10 5 cells) were harvested and seeded into Transwell inserts. VEGF-A (20 ng/ml) was added to the lower well of a Boyden chamber. (A) Representative photomicrographs of the HUVECs in the lower well of the Boyden chamber stained with crystal violet. Magnification ×100. (B) Cell migration of the HUVECs. Cell migration was quantified by counting the number of migrated cells and expressed as a percentage of the cell migration in the control. Three independent experiments were performed. * P
    Vegf A, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effect of curcumin and hypoxia on IGF-1R, p-Akt, p-Erk1/2, HIF-1α, VEGF protein expression in IGF-1R knockout HepG2 cells. (A) IGF-1R, p-Akt, p-Erk1/2, HIF-1α, VEGF protein expression were detected by western blot analysis. Graphic representation of relative density of (B) IGF-1R, (C) p-Akt, (D) p-Erk1/2, (E) HIF-1α and (F) VEGF protein levels, which were normalized to those of β-actin, Akt or Erk1/2, respectively. *P

    Journal: Experimental and Therapeutic Medicine

    Article Title: Effect of curcumin on vascular endothelial growth factor in hypoxic HepG2 cells via the insulin-like growth factor 1 receptor signaling pathway

    doi: 10.3892/etm.2018.5783

    Figure Lengend Snippet: Effect of curcumin and hypoxia on IGF-1R, p-Akt, p-Erk1/2, HIF-1α, VEGF protein expression in IGF-1R knockout HepG2 cells. (A) IGF-1R, p-Akt, p-Erk1/2, HIF-1α, VEGF protein expression were detected by western blot analysis. Graphic representation of relative density of (B) IGF-1R, (C) p-Akt, (D) p-Erk1/2, (E) HIF-1α and (F) VEGF protein levels, which were normalized to those of β-actin, Akt or Erk1/2, respectively. *P

    Article Snippet: The primary antibodies used in the experiment were as follows: Rabbit anti-IGF-1R (cat. no. 9750), rabbit anti-HIF-1α (cat. no. 3716), rabbit anti-VEGF (cat. no. 2463), rabbit anti-Akt (cat. no. 9272), rabbit anti-phosphorylated (p)-Akt (cat. no. 5012), rabbit anti-extracellular signal-regulated kinases (Erk1/2; cat. no. 4695), rabbit anti-p-Erk1/2 (cat. no. 4377) (all 1:1,000; Cell Signaling Technology, Inc., Danvers, MA, USA) and rabbit anti-β-actin (cat. no. sc-7210; 1:4,000; Santa Cruz Biotechnology, Inc.).

    Techniques: Expressing, Knock-Out, Western Blot

    Molecular mechanisms by which curcumin regulates the IGF-1R signaling pathway and the expression of VEGF in HepG2 cells under CoCl 2 -induced hypoxia. The arrow and blocked arrow (no arrowhead) indicate stimulation and inhibition, respectively. The dotted arrow indicates the translocation of HIF-1α and HIF-1β. The dotted blocked arrow indicates the potential effects of curcumin not directly examined in the present study. It was concluded that curcumin may suppress the expression of HIF-1 and VEGF by targeting IGF-1R or its downstream signaling pathways. VEGF, vascular endothelial growth factor; HIF-1α, hypoxia-inducible factor-1α; HIF-1β, hypoxia-inducible factor-1β; IGF-1, insulin-like growth factor-1; IGF-1R, insulin-like growth factor-1 receptor; PI3K, phosphoinositide-3-kinase; Akt, protein kinase B; MEK, mitogen-activated protein kinase-Erk kinase; Erk, extracellular signal-regulated kinase.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Effect of curcumin on vascular endothelial growth factor in hypoxic HepG2 cells via the insulin-like growth factor 1 receptor signaling pathway

    doi: 10.3892/etm.2018.5783

    Figure Lengend Snippet: Molecular mechanisms by which curcumin regulates the IGF-1R signaling pathway and the expression of VEGF in HepG2 cells under CoCl 2 -induced hypoxia. The arrow and blocked arrow (no arrowhead) indicate stimulation and inhibition, respectively. The dotted arrow indicates the translocation of HIF-1α and HIF-1β. The dotted blocked arrow indicates the potential effects of curcumin not directly examined in the present study. It was concluded that curcumin may suppress the expression of HIF-1 and VEGF by targeting IGF-1R or its downstream signaling pathways. VEGF, vascular endothelial growth factor; HIF-1α, hypoxia-inducible factor-1α; HIF-1β, hypoxia-inducible factor-1β; IGF-1, insulin-like growth factor-1; IGF-1R, insulin-like growth factor-1 receptor; PI3K, phosphoinositide-3-kinase; Akt, protein kinase B; MEK, mitogen-activated protein kinase-Erk kinase; Erk, extracellular signal-regulated kinase.

    Article Snippet: The primary antibodies used in the experiment were as follows: Rabbit anti-IGF-1R (cat. no. 9750), rabbit anti-HIF-1α (cat. no. 3716), rabbit anti-VEGF (cat. no. 2463), rabbit anti-Akt (cat. no. 9272), rabbit anti-phosphorylated (p)-Akt (cat. no. 5012), rabbit anti-extracellular signal-regulated kinases (Erk1/2; cat. no. 4695), rabbit anti-p-Erk1/2 (cat. no. 4377) (all 1:1,000; Cell Signaling Technology, Inc., Danvers, MA, USA) and rabbit anti-β-actin (cat. no. sc-7210; 1:4,000; Santa Cruz Biotechnology, Inc.).

    Techniques: Expressing, Inhibition, Translocation Assay

    Effect of CoCl 2 -induced hypoxia on IGF-1R, HIF-1α and VEGF protein expression in HepG2 cells. (A) Serum-starved IGF-1R knockout HepG2 cells were treated with different concentrations of CoCl 2 (0, 50, 100, 150, 200 and 400 µM) for 6 h. IGF-1R, HIF-1α and VEGF protein expression were detected by western blot analysis. (B) Western blot analysis data was quantified and the protein expression of IGF-1R, HIF-1α and VEGF are presented as a bar graph. *P

    Journal: Experimental and Therapeutic Medicine

    Article Title: Effect of curcumin on vascular endothelial growth factor in hypoxic HepG2 cells via the insulin-like growth factor 1 receptor signaling pathway

    doi: 10.3892/etm.2018.5783

    Figure Lengend Snippet: Effect of CoCl 2 -induced hypoxia on IGF-1R, HIF-1α and VEGF protein expression in HepG2 cells. (A) Serum-starved IGF-1R knockout HepG2 cells were treated with different concentrations of CoCl 2 (0, 50, 100, 150, 200 and 400 µM) for 6 h. IGF-1R, HIF-1α and VEGF protein expression were detected by western blot analysis. (B) Western blot analysis data was quantified and the protein expression of IGF-1R, HIF-1α and VEGF are presented as a bar graph. *P

    Article Snippet: The primary antibodies used in the experiment were as follows: Rabbit anti-IGF-1R (cat. no. 9750), rabbit anti-HIF-1α (cat. no. 3716), rabbit anti-VEGF (cat. no. 2463), rabbit anti-Akt (cat. no. 9272), rabbit anti-phosphorylated (p)-Akt (cat. no. 5012), rabbit anti-extracellular signal-regulated kinases (Erk1/2; cat. no. 4695), rabbit anti-p-Erk1/2 (cat. no. 4377) (all 1:1,000; Cell Signaling Technology, Inc., Danvers, MA, USA) and rabbit anti-β-actin (cat. no. sc-7210; 1:4,000; Santa Cruz Biotechnology, Inc.).

    Techniques: Expressing, Knock-Out, Western Blot

    Isomangiferin inhibits breast tumor growth and vascular endothelial growth factor receptor 2 (VEGFR-2) signaling pathway in vivo . Human breast cancer cells MDA-MB-231 were injected subcutanously into 5-week-old BALB/cA nude mice (2×10 6 per mouse). When subcutanous tumors grew to about 100 mm 3 , the mice were intraperitoneally treated with or without isomangiferin (10 mg/kg/day). (A) Photos for isomangiferin treated or non-treated tumors. (B) Isomangiferin supressed MDA-MB-231 xenografts growth. Tumor volume was recorded every 6 days and the tumor growth curve was drafted through Graphpad Prism 5 software package. Values are shown as mean±standard error of the mean (SEM) of three independent experiments. (C) Isomangiferin inhibited breast tumor growth as measured by tumor weight. Values are shown as mean±SEM of three independent experiments. (D) Immunohistochemical staining revealed that isomangiferin inhibited VEGFR-2 signaling pathway by blotting phosphorylated VEGFR-2 (p-VEGFR-2) and reducing CD31 expression. Tumor sections from isomangiferin-treated and isomangiferin-untreated groups were stained using p-VEGFR-2 and CD31 antibodies, and the number of positive cells was counted (IHC stain for p-VEGFR-2, CD31, ×400). * p

    Journal: Journal of Breast Cancer

    Article Title: Isomangiferin, a Novel Potent Vascular Endothelial Growth Factor Receptor 2 Kinase Inhibitor, Suppresses Breast Cancer Growth, Metastasis and Angiogenesis

    doi: 10.4048/jbc.2018.21.1.11

    Figure Lengend Snippet: Isomangiferin inhibits breast tumor growth and vascular endothelial growth factor receptor 2 (VEGFR-2) signaling pathway in vivo . Human breast cancer cells MDA-MB-231 were injected subcutanously into 5-week-old BALB/cA nude mice (2×10 6 per mouse). When subcutanous tumors grew to about 100 mm 3 , the mice were intraperitoneally treated with or without isomangiferin (10 mg/kg/day). (A) Photos for isomangiferin treated or non-treated tumors. (B) Isomangiferin supressed MDA-MB-231 xenografts growth. Tumor volume was recorded every 6 days and the tumor growth curve was drafted through Graphpad Prism 5 software package. Values are shown as mean±standard error of the mean (SEM) of three independent experiments. (C) Isomangiferin inhibited breast tumor growth as measured by tumor weight. Values are shown as mean±SEM of three independent experiments. (D) Immunohistochemical staining revealed that isomangiferin inhibited VEGFR-2 signaling pathway by blotting phosphorylated VEGFR-2 (p-VEGFR-2) and reducing CD31 expression. Tumor sections from isomangiferin-treated and isomangiferin-untreated groups were stained using p-VEGFR-2 and CD31 antibodies, and the number of positive cells was counted (IHC stain for p-VEGFR-2, CD31, ×400). * p

    Article Snippet: VEGFR-2 kinase activity (CST HTScan®, Cat No. #7788) was performed using a VEGFR-2 kinase assay kit purchased from Cell Signaling Technology.

    Techniques: In Vivo, Multiple Displacement Amplification, Injection, Mouse Assay, Software, Immunohistochemistry, Staining, Expressing

    Isomangiferin suppresses breast cancer through inhibiting the vascular endothelial growth factor receptor 2 (VEGFR-2) signaling pathway. (A) Isomangiferin induced cell apoptosis. Proteins from MDA-MB-231 cells treated with indicated concentrations of isomangiferin for 24 hours were submitted to Western blot for the immunoblotting of caspase-3 and cleaved poly ADP-ribose polymerase (cleaved PARP). (B) Isomangiferin's ihibiton on breast cancer cell proliferation was dependent on VEGFR-2 activity. 100 nM SU5408 was used or not to block VEGFR-2's activity and then MDA-MB-231 cells were treated with isomangiferin. The cell proliferation was assessed by MTS assay. Values are shown as mean±standard error of the mean of three independent experiments. (C) Isomangiferin suppressed the activation of VEGFR-2 in human umbilical vein endothelial cells (HUVECs). The activation of VEGFR-2 was analyzed by Western blot and probed with indicated antibodies. Western blot was conducted in the way that described in Methods and specific antibodies were used accordingly. PARP=poly ADP-ribose polymerase; NS=not significant; p-AKT=phosphorylated protein kinase B, p-PKB/p-AKT; AKT=protein kinase B, PKB/AKT; p-ERK=phosphorylated extracellular regulated protein kinases; ERK=extracellular regulated protein kinases; p-STAT3=phosphorylated signal transducer and activator of transcription 3; FAK=focal adhesion kinase. * p

    Journal: Journal of Breast Cancer

    Article Title: Isomangiferin, a Novel Potent Vascular Endothelial Growth Factor Receptor 2 Kinase Inhibitor, Suppresses Breast Cancer Growth, Metastasis and Angiogenesis

    doi: 10.4048/jbc.2018.21.1.11

    Figure Lengend Snippet: Isomangiferin suppresses breast cancer through inhibiting the vascular endothelial growth factor receptor 2 (VEGFR-2) signaling pathway. (A) Isomangiferin induced cell apoptosis. Proteins from MDA-MB-231 cells treated with indicated concentrations of isomangiferin for 24 hours were submitted to Western blot for the immunoblotting of caspase-3 and cleaved poly ADP-ribose polymerase (cleaved PARP). (B) Isomangiferin's ihibiton on breast cancer cell proliferation was dependent on VEGFR-2 activity. 100 nM SU5408 was used or not to block VEGFR-2's activity and then MDA-MB-231 cells were treated with isomangiferin. The cell proliferation was assessed by MTS assay. Values are shown as mean±standard error of the mean of three independent experiments. (C) Isomangiferin suppressed the activation of VEGFR-2 in human umbilical vein endothelial cells (HUVECs). The activation of VEGFR-2 was analyzed by Western blot and probed with indicated antibodies. Western blot was conducted in the way that described in Methods and specific antibodies were used accordingly. PARP=poly ADP-ribose polymerase; NS=not significant; p-AKT=phosphorylated protein kinase B, p-PKB/p-AKT; AKT=protein kinase B, PKB/AKT; p-ERK=phosphorylated extracellular regulated protein kinases; ERK=extracellular regulated protein kinases; p-STAT3=phosphorylated signal transducer and activator of transcription 3; FAK=focal adhesion kinase. * p

    Article Snippet: VEGFR-2 kinase activity (CST HTScan®, Cat No. #7788) was performed using a VEGFR-2 kinase assay kit purchased from Cell Signaling Technology.

    Techniques: Multiple Displacement Amplification, Western Blot, Activity Assay, Blocking Assay, MTS Assay, Activation Assay

    Knockdown of paxillin inhibits the VEGF-A-induced migration of HUVECs. Cells were cultured on plates following either mock-transfection or transfection with scramble siRNA or siPXN-1, respectively. HUVECs (2×10 5 cells) were harvested and seeded into Transwell inserts. VEGF-A (20 ng/ml) was added to the lower well of a Boyden chamber. (A) Representative photomicrographs of the HUVECs in the lower well of the Boyden chamber stained with crystal violet. Magnification ×100. (B) Cell migration of the HUVECs. Cell migration was quantified by counting the number of migrated cells and expressed as a percentage of the cell migration in the control. Three independent experiments were performed. * P

    Journal: Molecular Medicine Reports

    Article Title: Paxillin regulates vascular endothelial growth factor A-induced in vitro angiogenesis of human umbilical vein endothelial cells

    doi: 10.3892/mmr.2014.2961

    Figure Lengend Snippet: Knockdown of paxillin inhibits the VEGF-A-induced migration of HUVECs. Cells were cultured on plates following either mock-transfection or transfection with scramble siRNA or siPXN-1, respectively. HUVECs (2×10 5 cells) were harvested and seeded into Transwell inserts. VEGF-A (20 ng/ml) was added to the lower well of a Boyden chamber. (A) Representative photomicrographs of the HUVECs in the lower well of the Boyden chamber stained with crystal violet. Magnification ×100. (B) Cell migration of the HUVECs. Cell migration was quantified by counting the number of migrated cells and expressed as a percentage of the cell migration in the control. Three independent experiments were performed. * P

    Article Snippet: Immunoprecipitation The HUVECs were grown to confluence and stimulated with 20 ng/ml VEGF-A (Cell Signaling Technology, Inc., Beverly, MA, USA) at 37°C for 0, 20, 40 and 60 min.

    Techniques: Migration, Cell Culture, Transfection, Staining

    Knock down of paxillin inhibits the VEGF-A-induced proliferation of HUVECs. The HUVECs were cultured on plates following either mock-transfected or transfected with scramble siRNA or siPXN-1, respectively. VEGF-A (20 ng/ml) was added to the medium. (A) Representative photomicrographs of the HUVECs stained with EdU (red) and Hoechst 33342 (blue). Magnification ×200. (B) Percentage of EdU-positive cells. * P

    Journal: Molecular Medicine Reports

    Article Title: Paxillin regulates vascular endothelial growth factor A-induced in vitro angiogenesis of human umbilical vein endothelial cells

    doi: 10.3892/mmr.2014.2961

    Figure Lengend Snippet: Knock down of paxillin inhibits the VEGF-A-induced proliferation of HUVECs. The HUVECs were cultured on plates following either mock-transfected or transfected with scramble siRNA or siPXN-1, respectively. VEGF-A (20 ng/ml) was added to the medium. (A) Representative photomicrographs of the HUVECs stained with EdU (red) and Hoechst 33342 (blue). Magnification ×200. (B) Percentage of EdU-positive cells. * P

    Article Snippet: Immunoprecipitation The HUVECs were grown to confluence and stimulated with 20 ng/ml VEGF-A (Cell Signaling Technology, Inc., Beverly, MA, USA) at 37°C for 0, 20, 40 and 60 min.

    Techniques: Cell Culture, Transfection, Staining

    Knockdown of paxillin inhibits the VEGF-A-induced adhesion of HUVECs. The HUVECs were seeded into 96-well plates following either mock-transfection or transfection with scramble siRNA or siPXN-1, respectively. VEGF-A (20 ng/ml) was added to the medium. (A) Representative photomicrographs of the HUVECs following siRNA and VEGF-A transfection. Magnification ×200. (B) OD at a wavelength of 490 nm of the HUVECs stained with MTT. Three independent experiments were performed. * P

    Journal: Molecular Medicine Reports

    Article Title: Paxillin regulates vascular endothelial growth factor A-induced in vitro angiogenesis of human umbilical vein endothelial cells

    doi: 10.3892/mmr.2014.2961

    Figure Lengend Snippet: Knockdown of paxillin inhibits the VEGF-A-induced adhesion of HUVECs. The HUVECs were seeded into 96-well plates following either mock-transfection or transfection with scramble siRNA or siPXN-1, respectively. VEGF-A (20 ng/ml) was added to the medium. (A) Representative photomicrographs of the HUVECs following siRNA and VEGF-A transfection. Magnification ×200. (B) OD at a wavelength of 490 nm of the HUVECs stained with MTT. Three independent experiments were performed. * P

    Article Snippet: Immunoprecipitation The HUVECs were grown to confluence and stimulated with 20 ng/ml VEGF-A (Cell Signaling Technology, Inc., Beverly, MA, USA) at 37°C for 0, 20, 40 and 60 min.

    Techniques: Transfection, Staining, MTT Assay

    Tube formation of HUVECs is inhibited by siRNA-mediated paxillin knockdown. The HUVECs were seeded onto Matrigel 48 h after being either mock-transfected or transfected with scramble siRNA or siPXN-1, respectively. VEGF-A (20 ng/ml) was added to the medium. (A) Photomicrographs of the tubes; (B) Tube length was calculated using the Image-Pro Plus image processing system. Three independent experiments were performed. * P

    Journal: Molecular Medicine Reports

    Article Title: Paxillin regulates vascular endothelial growth factor A-induced in vitro angiogenesis of human umbilical vein endothelial cells

    doi: 10.3892/mmr.2014.2961

    Figure Lengend Snippet: Tube formation of HUVECs is inhibited by siRNA-mediated paxillin knockdown. The HUVECs were seeded onto Matrigel 48 h after being either mock-transfected or transfected with scramble siRNA or siPXN-1, respectively. VEGF-A (20 ng/ml) was added to the medium. (A) Photomicrographs of the tubes; (B) Tube length was calculated using the Image-Pro Plus image processing system. Three independent experiments were performed. * P

    Article Snippet: Immunoprecipitation The HUVECs were grown to confluence and stimulated with 20 ng/ml VEGF-A (Cell Signaling Technology, Inc., Beverly, MA, USA) at 37°C for 0, 20, 40 and 60 min.

    Techniques: Transfection

    VEGF-A induces the phosphorylation of paxillin in HUVECs. The HUVECs were treated with 20 ng/ml VEGF-A for 0, 20, 40 and 60 min. Western blot analysis was performed to assess the phosphorylation of paxillin phosphorylated at PY118 and PY31 as well as total paxillin protein. Actin was used as a loading control. VEGF-A, vascular endothelial growth factor A; HUVECs, human umbilical vein endothelial cells. PY118, tyrosine 118; PY31, tyrosine31.

    Journal: Molecular Medicine Reports

    Article Title: Paxillin regulates vascular endothelial growth factor A-induced in vitro angiogenesis of human umbilical vein endothelial cells

    doi: 10.3892/mmr.2014.2961

    Figure Lengend Snippet: VEGF-A induces the phosphorylation of paxillin in HUVECs. The HUVECs were treated with 20 ng/ml VEGF-A for 0, 20, 40 and 60 min. Western blot analysis was performed to assess the phosphorylation of paxillin phosphorylated at PY118 and PY31 as well as total paxillin protein. Actin was used as a loading control. VEGF-A, vascular endothelial growth factor A; HUVECs, human umbilical vein endothelial cells. PY118, tyrosine 118; PY31, tyrosine31.

    Article Snippet: Immunoprecipitation The HUVECs were grown to confluence and stimulated with 20 ng/ml VEGF-A (Cell Signaling Technology, Inc., Beverly, MA, USA) at 37°C for 0, 20, 40 and 60 min.

    Techniques: Western Blot