Journal: The Scientific World Journal

Article Title: Biological and Demographic Profile of Meningiomas in a Cohort of Egyptian Patients: Impact on Tumor Recurrence

doi: 10.1155/2013/375139

Figure Lengend Snippet: (a) EMA immunostain highlighting whorls of meningioma cells (200x); (b) intense and diffuse VEGF expression in an anaplastic meningioma (200x); (c) Ki67 immunostain revealing a moderate proliferative activity in a case of atypical meningioma (200x); (d) benign meningioma with significant PR expression (200x); (e) CD20 highlighting rare B lymphocytes (100x); (f) CD3 showing a large population of T lymphocytes (100x).

Article Snippet: The used primary antibodies (at 1 : 100 dilution): vascular endothelial growth factor (VEGF), clone: VG; Ki67, clone: SP6; progesterone receptor (PR), clone: SP2; CD20 Ab-1, clone: L26; and CD3epsilon Ab-2, clone: PS1, as well as the detection kit (UltraVision Detection System Anti-Polyvalent, HRP/DAB, Ready-To-Use), were purchased from Thermo Scientific Lab Vision, USA.

Techniques: Expressing, Activity Assay

Journal: Scientific Reports

Article Title: Differential response of rat strains to obesogenic diets underlines the importance of genetic makeup of an individual towards obesity

doi: 10.1038/s41598-017-09149-6

Figure Lengend Snippet: Effect of high calorie environment towards oxidative stress and pro-inflammatory status in rat strains. After the completion of feeding experiment, blood was collected; plasma was separated and estimated the levels of pro-inflammatory cytokines. ( A ) Interleukin 6; IL-6. ( B ) Tumor necrosis factor alpha; TNFα. ( C ) Interleukin 1 beta; IL-1β. ( D ) Macrophage inflammatory protein 1 alpha; MIP-1α. ( E ) Vascular endothelial growth factor; VEGF. ( F ) IFN gamma inducible protein 10; IP-10. ( G ) Monocyte chemotactic protein 1; MCP1 and anti -inflammatory cytokines such as ( H ) Interleukin 4; IL-4 and ( I ) Interleukin 10; IL-10. ( J ) TBARS assay was performed on liver tissue lysate to study oxidative stress. Data was presented as mean ± SEM (n = 6 per group). Diets: control; HF, high fat; HS, high sucrose; HFS, high fat sucrose. TBARS, Thio Barbituricacid Reactive Substances. *P

Article Snippet: Rat Cytokine/Chemokine Profile The adipocytokines such as leptin, macrophage inflammatory protein-1 alpha (MIP-1α, CCL3), interleukin-4 (IL-4), interleukin-1 beta (IL-1β), interleukin-6 (IL-6), interleukin-10 (IL-10), macrophage chemo attractant protein-1 (MCP-1, CCL2), IFN-gamma-inducible protein 10 (IP- 10, CXCL10), vascular endothelial growth factor (VEGF), and tumor necrosis factor-alpha (TNF-α) were estimated both in circulation as well as in visceral adipose tissue by Milliplex Rat cytokine immunoassay kit (Millipore, USA) according to manufacturer’s instructions.

Techniques: TBARS Assay

Journal: The Journal of Biological Chemistry

Article Title: Bone Morphogenetic Protein Receptor II Is a Novel Mediator of Endothelial Nitric-oxide Synthase Activation *

doi: 10.1074/jbc.M111.274100

Figure Lengend Snippet: BMP2 and BMP4 stimulate eNOS phosphorylation in PAECs. A and B , representative Western blots of bPAECs stimulated with BMP2 ( A ) or BMP4 ( B ) for various times and analyzed for eNOS phosphorylation at serine 1179. VEGF was used as a positive control. C

Article Snippet: BMP4 and vascular endothelial growth factor (VEGF) were from R & D Systems (Minneapolis, MN).

Techniques: Western Blot, Positive Control

Journal: The Journal of Biological Chemistry

Article Title: Bone Morphogenetic Protein Receptor II Is a Novel Mediator of Endothelial Nitric-oxide Synthase Activation *

doi: 10.1074/jbc.M111.274100

Figure Lengend Snippet: BMP2 and BMP4 stimulate eNOS enzymatic activity in PAECs. bPAECs were stimulated with BMP2, BMP4, or VEGF for 30 min, and lysates were analyzed for eNOS activity by measuring the conversion of arginine to citrulline. l -NAME added to the enzyme activity

Article Snippet: BMP4 and vascular endothelial growth factor (VEGF) were from R & D Systems (Minneapolis, MN).

Techniques: Activity Assay

Journal: The Journal of Biological Chemistry

Article Title: Bone Morphogenetic Protein Receptor II Is a Novel Mediator of Endothelial Nitric-oxide Synthase Activation *

doi: 10.1074/jbc.M111.274100

Figure Lengend Snippet: BMP2- and BMP4-stimulated eNOS phosphorylation is dependent on BMPRII. Serum-starved hPAECs transfected with control or BMPRII siRNA were stimulated for various times with BMP2, BMP4, or VEGF and then assayed for eNOS phosphorylation by Western blot analysis

Article Snippet: BMP4 and vascular endothelial growth factor (VEGF) were from R & D Systems (Minneapolis, MN).

Techniques: Transfection, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Bone Morphogenetic Protein Receptor II Is a Novel Mediator of Endothelial Nitric-oxide Synthase Activation *

doi: 10.1074/jbc.M111.274100

Figure Lengend Snippet: BMP2 and BMP4 stimulate eNOS-dependent PAEC migration. PAECs were stimulated with VEGF, BMP2, or BMP4 with or without l -NAME (100 μ m ) and assayed for migration using the monolayer wound assay. Data represent the mean ± S.D. ( error bars

Article Snippet: BMP4 and vascular endothelial growth factor (VEGF) were from R & D Systems (Minneapolis, MN).

Techniques: Migration

Journal: The Journal of Biological Chemistry

Article Title: Bone Morphogenetic Protein Receptor II Is a Novel Mediator of Endothelial Nitric-oxide Synthase Activation *

doi: 10.1074/jbc.M111.274100

Figure Lengend Snippet: BMP2 and BMP4 elicit canonical changes in eNOS protein-protein interactions. eNOS was immunoprecipitated ( IP ) from bPAECs stimulated for 30 min with BMP2, BMP4, or VEGF. Immunoprecipitates were subjected to SDS-PAGE and analyzed by Western blotting for

Article Snippet: BMP4 and vascular endothelial growth factor (VEGF) were from R & D Systems (Minneapolis, MN).

Techniques: Immunoprecipitation, SDS Page, Western Blot

Journal: NPJ Breast Cancer

Article Title: Activin-A signaling promotes epithelial–mesenchymal transition, invasion, and metastatic growth of breast cancer

doi: 10.1038/npjbcancer.2015.7

Figure Lengend Snippet: Activin promotes tumorigenicity of breast cancer cells in immunocompromised mice. ( a ) Stable overexpression of activin-A in MCF-7 enhances (i), whereas stable knockdown of activin-A in MDA-MB-231 cells (ii) reduces their tumor-forming ability in nude mice. Shown below are the representative images of the tumors formed s.c. ( b ) Immunohistochemical analysis of MCF-7-overexpressing tumors shows EMT-like changes and higher ki-67 index. ( c ) Treatment of MCF-7 cells with activin-A (i) or overexpression of activin-A in MCF-7 cells (ii) results in increased levels of vascular endothelial growth factor-A (VEGF-A). ( d ) Luciferase reporter assay in HEK 293T cells shows that activin-A regulates VEGF promoter activity. ( e ) Tail vein injection of activin-A-overexpressing MCF-7 cells shows better tumor-forming ability in the livers of nude mice (i). Normal is shown here as the reference from an animal without injection of any cells. The panel below (ii) shows the hematoxylin and eosin (H E) staining of the liver tissue sections. The graph (iii) and (iv) shows number and size of nodules formed in the liver per animal. ( f ) CD44 high/ CD24 low fluorescence-activated cell sorting (FACS) analysis of activin-A-overexpressing or knockdown cells shows that activin-A influences stemness of breast cancer cells (i). Quantitative PCR analysis shows that treatment of MCF-7 or MDA-MB-231 cells with recombinant activin-A induces various markers of stemness (ii).

Article Snippet: Reagents Recombinant human activin-A (338-AC-010) and activin-A antibody (AF338) were purchased from R & D Systems (Minneapolis, MN, USA); phosphoSMAD2 (3101 and 9510) from Cell Signaling Technology (Boston, MA, USA); SMAD3 (1735), E-cadherin (1702), N-cadherin (2019), and α-smooth muscle actin (1184-1) from Epitomics (CA, USA); Vimentin (V2258) and fluorescein isothiocyanate-conjugated phalloidin (P5282) from Sigma (St Louis, MO, USA); phosphoSMAD3 (ab52903) and BMP2 (ab14933) from Abcam (Cambridge, MA, USA); vascular endothelial growth factor-A (VEGF-A) (M7273) from Dako (Denmark); and PE-CD44 (560533)/PE-cy7 CD24 (555428) from BD (NJ, USA).

Techniques: Mouse Assay, Over Expression, Multiple Displacement Amplification, Immunohistochemistry, Luciferase, Reporter Assay, Activity Assay, Injection, Staining, Fluorescence, FACS, Real-time Polymerase Chain Reaction, Recombinant

Journal: Oncology Letters

Article Title: Effect of cycloxygenase-2 silencing on the malignant biological behavior of MCF-7 breast cancer cells

doi: 10.3892/ol.2014.2395

Figure Lengend Snippet: VEGF-A and -C protein expression of breast cancer MCF-7 cells in the COX-2-shRNA group was significantly lower than that of the blank and mock groups. COX-2, cyclooxygenase-2; VEGF, vascular endothelial growth factor; shRNA, short hairpin RNA.

Article Snippet: 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.

Techniques: Expressing, shRNA

Journal: mAbs

Article Title: VEGF neutralizing aerosol therapy in primary pulmonary adenocarcinoma with K-ras activating-mutations

doi: 10.4161/mabs.34454

Figure Lengend Snippet: Anti-VEGF mAb impairs tumor cell proliferation

Article Snippet: Native or aerosolized G6–31 at 1, 5 or 10 nM in 1X PBS was pre-incubated with 0.5 nM of human VEGF-A (Cell Signaling) for 60 min, and then added to the cells.

Techniques:

Journal: mAbs

Article Title: VEGF neutralizing aerosol therapy in primary pulmonary adenocarcinoma with K-ras activating-mutations

doi: 10.4161/mabs.34454

Figure Lengend Snippet: VEGF neutralization impairs VEGFR2 activation

Article Snippet: Native or aerosolized G6–31 at 1, 5 or 10 nM in 1X PBS was pre-incubated with 0.5 nM of human VEGF-A (Cell Signaling) for 60 min, and then added to the cells.

Techniques: Neutralization, Activation Assay

Journal: mAbs

Article Title: VEGF neutralizing aerosol therapy in primary pulmonary adenocarcinoma with K-ras activating-mutations

doi: 10.4161/mabs.34454

Figure Lengend Snippet: Anti-VEGF mAb inhibits angiogenesis

Article Snippet: Native or aerosolized G6–31 at 1, 5 or 10 nM in 1X PBS was pre-incubated with 0.5 nM of human VEGF-A (Cell Signaling) for 60 min, and then added to the cells.

Techniques:

Journal: mAbs

Article Title: VEGF neutralizing aerosol therapy in primary pulmonary adenocarcinoma with K-ras activating-mutations

doi: 10.4161/mabs.34454

Figure Lengend Snippet: Aerosolized anti-VEGF mAb passes slowly and poorly into the systemic circulation

Article Snippet: Native or aerosolized G6–31 at 1, 5 or 10 nM in 1X PBS was pre-incubated with 0.5 nM of human VEGF-A (Cell Signaling) for 60 min, and then added to the cells.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Enhanced Therapeutic Potential of the Secretome Released from Adipose-Derived Stem Cells by PGC-1α-Driven Upregulation of Mitochondrial Proliferation

doi: 10.3390/ijms20225589

Figure Lengend Snippet: In vivo effects of PGC-secretome on liver regeneration in partially hepatectomized mice. ( A ) (Right) Western blot analysis of the liver at the 7th day following each treatment. The markers studied in the western blot analysis included those for liver regeneration (PCNA, HGF, VEGF, p-STAT3, and t-STAT3) and anti-apoptosis (Bcl-xL). The PGC-secretome group displayed a significant increase in the markers for liver regeneration and anti-apoptosis compared to the levels in the normal secretome group. (Left and Below) Relative densities of the markers in each group. ( B ) Liver regeneration rate (%) based on the ratio of liver weight to body weight (LW/BW) on days 2 and 7 after 70% partial hepatectomy. The PGC-secretome group displayed the highest LW/BW ratios on day 7 after partial hepatectomy. Values are presented as mean ± standard deviation of three independent experiments; * p

Article Snippet: Primary antibodies against proliferating cell nuclear antigen (PCNA), phosphorylated-signal transducer and activator of transcription 3 (p-STAT3), STAT3, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), B-cell leukemia-extra large (Bcl-xL), dynamin related protein 1 (DRP-1), and β-actin were all obtained from Cell Signaling Technology (Beverly, MA, USA), and Opa1 mitochondrial dynamin like GTPase (OPA-1) was obtained from Santa Cruz biotechnology (Santa Cruz, CA, USA).

Techniques: In Vivo, Pyrolysis Gas Chromatography, Mouse Assay, Western Blot, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Enhanced Therapeutic Potential of the Secretome Released from Adipose-Derived Stem Cells by PGC-1α-Driven Upregulation of Mitochondrial Proliferation

doi: 10.3390/ijms20225589

Figure Lengend Snippet: In vitro validation of the effects of the peroxisome proliferator activated receptor λ coactivator 1α (PGC-secretome) on cell viability and expression of various markers. ( A ) Effects of each secretome on the viability of AML12 hepatocytes. Cell viability analysis revealed that the PGC-secretome increased the viability of thioacetamide (TAA)-treated AML12 hepatocytes significantly more than did the control and normal secretome. ( B ) Western blot analysis demonstrating the effects of each secretome on the expression of various markers in AML12 hepatocytes. The markers included those for liver proliferation (p-STAT3, t-STAT3, VEGF, and HGF), mitochondrial fusion (OPA-1), mitochondrial fission (DRP-1), pro-apoptosis (BIM), and anti-apoptosis (Bcl-xL). PGC-secretome significantly increased the expression of markers related to proliferation, mitochondrial fusion, and anti-apoptosis, and significantly decreased the expression of the markers related to mitochondrial fission and pro-apoptosis. Values are presented as mean ± standard deviation of three independent experiments; * p

Article Snippet: Primary antibodies against proliferating cell nuclear antigen (PCNA), phosphorylated-signal transducer and activator of transcription 3 (p-STAT3), STAT3, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), B-cell leukemia-extra large (Bcl-xL), dynamin related protein 1 (DRP-1), and β-actin were all obtained from Cell Signaling Technology (Beverly, MA, USA), and Opa1 mitochondrial dynamin like GTPase (OPA-1) was obtained from Santa Cruz biotechnology (Santa Cruz, CA, USA).

Techniques: In Vitro, Pyrolysis Gas Chromatography, Expressing, Western Blot, Standard Deviation

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

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

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

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

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

Journal: British Journal of Cancer

Article Title: Loss of Smad4 in colorectal cancer induces resistance to 5-fluorouracil through activating Akt pathway

doi: 10.1038/bjc.2013.789

Figure Lengend Snippet: Smad4 downregulates VEGF and inhibits angiogenesis. ( A ) VEGF expression in CT26 and SW620 cell clones was examined by western blot analyses. ( B ) Representative tumour masses with or without 5-FU treatment are shown. Visible vessels are marked by black arrows. Formalin-fixed paraffin-embedded tissue sections from ( C ) orthotopic tumours and ( D ) subcutaneous tumours were subjected to H E and CD31 staining. Representative pictures are shown ( × 100 and × 400) with vessel density and vessel number. * P

Article Snippet: Antibodies were purchased as follows: Santa Cruz Biotechnology (Santa Cruz, CA, USA): anti-Smad4, anti-p21Cip1 , anti-p27Kip1 , anti-Cyclin D1, anti-Survivin, anti-Bcl-2, anti-VEGF; Cell Signaling (Denver, MA, USA): anti-PARP, anti-cleaved-Caspase3, anti-p-Akt, anti-Akt, anti-Bcl-w, anti-Bcl-xL, anti-Bad, anti-Bim, anti-Bax, anti-PUMA; Zymed Laboratories Inc. (San Francisco, CA, USA): anti-c-Myc.

Techniques: Expressing, Clone Assay, Western Blot, Formalin-fixed Paraffin-Embedded, Staining

Journal: Molecular Medicine Reports

Article Title: CCL18 enhances migration, invasion and EMT by binding CCR8 in bladder cancer cells

doi: 10.3892/mmr.2018.9791

Figure Lengend Snippet: CCL18 affects the expression levels of E-cadherin, MMP-2 and VEGF-C. (A and B) 5637 bladder cancer cells were treated with PBS or CCL18 (50 or 100 ng/ml) for 36 h, with or without pretreatment with the CCR8 inhibitor R243. Protein expression levels of E-cadherin, MMP-2 and VEGF-C were detected in 5637 cells by western blot analysis. *P

Article Snippet: Once proteins were transferred to nitrocellulose membranes, they were incubated with antibodies targeting β-actin (cat. no. 58169, 1:1,000; Cell Signaling Technology, Inc., Danvers, MA, USA), CCR8 (cat. no. ab32131, 1:500; Abcam, Cambridge, UK), E-cadherin (cat. no. 3195, 1:1,000; Cell Signaling Technology, Inc., Danvers, MA, USA), matrix metalloproteinase (MMP)-2 (cat. no. 40994, 1:1,000; Cell Signaling Technology, Inc.) and vascular endothelial growth factor (VEGF)-C (cat. no. 2445, 1:1,000; Cell Signaling Technology, Inc.), followed by incubation with a horseradish peroxidase-labeled goat anti-rabbit secondary antibody (cat. no. ab6721, 1:3,000; Abcam) for 1.5 h at room temperature.

Techniques: Expressing, Western Blot

Journal: Molecular Medicine Reports

Article Title: CCL18 enhances migration, invasion and EMT by binding CCR8 in bladder cancer cells

doi: 10.3892/mmr.2018.9791

Figure Lengend Snippet: Effects of CCR8 shRNA on CCL18-induced expression of E-cadherin, MMP-2 and VEGF-C. 5637 bladder cancer cells were treated with CCL18 (50 ng/ml), CCL18 + NT shRNA or CCL18 + CCR8 shRNA for 36 h. (A) mRNA expression levels of CCR8 after being transfected with CCR8 shRNA was examined by reverse transcription-quantitative polymerase chain reaction. β-actin was used as an internal control. (B) mRNA expression levels of CCR8, E-cadherin, MMP-2 and VEGF-C were examined by reverse transcription-quantitative polymerase chain reaction. β-actin was used as an internal control. (C and D) Protein expression levels of CCR8, E-cadherin, MMP-2 and VEGF-C in 5637 cells were detected by western blotting. β-actin was used as an internal control. *P

Article Snippet: Once proteins were transferred to nitrocellulose membranes, they were incubated with antibodies targeting β-actin (cat. no. 58169, 1:1,000; Cell Signaling Technology, Inc., Danvers, MA, USA), CCR8 (cat. no. ab32131, 1:500; Abcam, Cambridge, UK), E-cadherin (cat. no. 3195, 1:1,000; Cell Signaling Technology, Inc., Danvers, MA, USA), matrix metalloproteinase (MMP)-2 (cat. no. 40994, 1:1,000; Cell Signaling Technology, Inc.) and vascular endothelial growth factor (VEGF)-C (cat. no. 2445, 1:1,000; Cell Signaling Technology, Inc.), followed by incubation with a horseradish peroxidase-labeled goat anti-rabbit secondary antibody (cat. no. ab6721, 1:3,000; Abcam) for 1.5 h at room temperature.

Techniques: shRNA, Expressing, Transfection, Real-time Polymerase Chain Reaction, Western Blot

Journal: PLoS ONE

Article Title: Activation of the Wnt/?-Catenin Signaling Pathway by Mechanical Ventilation Is Associated with Ventilator-Induced Pulmonary Fibrosis in Healthy Lungs

doi: 10.1371/journal.pone.0023914

Figure Lengend Snippet: Mechanical ventilation increased MMP-7, cyclin D1, and VEGF in the lung. Densitometry analysis of the active form (20 kDa) of MMP-7 was normalized to the inactive form (30 kDa) and then normalized to β-actin. Densitometry analysis (in graph) and representative gels correspond to three different experimental protocols: spontaneous breathing animals (controls, C), ventilated with low V T and ventilated with high V T . (*) p

Article Snippet: Detection of WNT5A, AXIN2 (Abcam, Cambridge, UK), total β-catenin, MMP-7, cyclin D1, and vascular endothelial growth factor (VEGF) (Santa Cruz Biotechnology, Santa Cruz, CA), non-phospho (Ser33/37/Thr41) β-catenin (Cell Signaling Technology, Massachusetts) were performed in random samples by Western blotting using rabbit polyclonal anti-WNT5A, anti-AXIN2, anti-β-catenin, anti-non-phospho (Ser33/37/Thr41) β-catenin, and anti-MMP-7 antibodies, and a goat anti-rabbit IgG-HRP as secondary antibody (Santa Cruz Biotechnology, Santa Cruz, CA), mouse monoclonal antibody anti-cyclin D1 and a rabbit anti-mouse IgG-HRP as secondary antibody (Dako, Glostrup, Denmark), goat polyclonal anti-VEFG and a donkey anti-goat IgG-HRP (Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques:

Journal: PLoS ONE

Article Title: Deletion of Thioredoxin Interacting Protein (TXNIP) Augments Hyperoxia-Induced Vaso-Obliteration in a Mouse Model of Oxygen Induced-Retinopathy

doi: 10.1371/journal.pone.0110388

Figure Lengend Snippet: Deletion of TXNIP does not alter VEGFR2/Akt activation under hyperoxia. Wild type (WT) and TXNIP knockout (TKO) mice were subjected to hyperoxia (75% O2, p7–p12). Activation of VEGFR2 ( A ) and Akt ( B ) were examined as downstream signal of VEGF in p12 WT and TKO retinas. TKO showed significant decrease in phosphorylation of VEGFR-2 and Akt compared to WT under normoxic condition. We did not detect significant change in the activation of VEGFR2 and its downstream Akt in retinas from TKO and WT in response to hyperoxia. (*P

Article Snippet: The primary antibodies were purchased as follow: VEGF (Rabbit polyclonal, EMD-Millipore), phosphor-VEGFR2, VEGFR2, phospho-Akt, Akt, phospho-ASK-1, ASK-1, cleaved caspase-3 (Rabbit polyclonal, Cell Signaling Tech, Danvers, MA), total Trx (Mouse monoclonal, Santa Cruz, Dallas, TX), and TXNIP (Rabbit polyclonal, Invitrogen, Grand Island, NY), cleaved PARP (BD Bioscience Pharmingen, San Diego, CA).

Techniques: Activation Assay, Knock-Out, Mouse Assay

Journal: PLoS ONE

Article Title: Deletion of Thioredoxin Interacting Protein (TXNIP) Augments Hyperoxia-Induced Vaso-Obliteration in a Mouse Model of Oxygen Induced-Retinopathy

doi: 10.1371/journal.pone.0110388

Figure Lengend Snippet: Representative diagram shows the impact of TXNIP deletion on retina vasculature under both normoxia and hyperoxia. Under normoxia, retinas from TXNIP-deficient mice showed similar VEGF levels, less peroxynitrite (ONOO-) levels, less VEGF receptor-2 (pVEGFR2) activation and upregulated thioredoxin (Trx) that collectively lead to normal vascular development in comparison to WT mice. Under hyperoxia, retinas from WT mice showed higher peroxynitrite formation, less survival Akt activation (pAkt) and upregulated proapoptotic signal of ASK-1 resulting in vaso-obliteration. Retinas from TKO although showed less peroxynitrite levels and maintained Akt activation, retinas experienced significant decreases in thioredoxin (Trx) that shift the balance of the ASK-1-Trx inhibitory complex and increases the activation of the proapoptotic ASK-1 pathway leading to exacerbated vasoobliteration compared to WT.

Article Snippet: The primary antibodies were purchased as follow: VEGF (Rabbit polyclonal, EMD-Millipore), phosphor-VEGFR2, VEGFR2, phospho-Akt, Akt, phospho-ASK-1, ASK-1, cleaved caspase-3 (Rabbit polyclonal, Cell Signaling Tech, Danvers, MA), total Trx (Mouse monoclonal, Santa Cruz, Dallas, TX), and TXNIP (Rabbit polyclonal, Invitrogen, Grand Island, NY), cleaved PARP (BD Bioscience Pharmingen, San Diego, CA).

Techniques: Mouse Assay, Activation Assay

Journal: Stem Cells Translational Medicine

Article Title: Enhanced Cardioprotection by Human Endometrium Mesenchymal Stem Cells Driven by Exosomal MicroRNA‐21

doi: 10.5966/sctm.2015-0386

Figure Lengend Snippet: Schematic of superior cardioprotection, including antiapoptosis and angiogenesis, mediated by exosomal miR‐2 1 secreted by EnMSCs. Abbreviations: Bax, Bcl‐2‐associated X protein; Bcl‐2, B‐cell CLL/lymphoma 2; Casp‐3, caspase‐3; EnMSCs, endometrium‐derived mesenchymal stem cells; miR‐21, microRNA‐21; p‐Akt, phosphorylated Akt; PTEN, phosphatase and tensin homolog; VEFG, vascular endothelial growth factor.

Article Snippet: After 1 hour of blocking with PBST containing 5% BSA at room temperature, primary antibodies, including anti‐Bcl‐2, Bcl‐2‐associated X protein (Bax), caspase‐3, phosphorylated‐Akt/total‐Akt, PTEN, vascular endothelial growth factor (VEGF), CD63, and β‐actin (Abcam) were diluted to 1:1,000 and incubated with the membrane overnight at 4°C.

Techniques: Derivative Assay

Journal: Oncotarget

Article Title: Prognostic significance of Cytokeratin 20-positive lymph node vascular endothelial growth factor A mRNA and chromodomain helicase DNA binding protein 4 in pN0 colorectal cancer patients

doi: 10.18632/oncotarget.23424

Figure Lengend Snippet: Kaplan-Meier curves of recurrence for the 2nd cohort of 47 CK20+ pN0 CRC patients after follow-up for 60 months based on VEGF-A mRNA copy numbers

Article Snippet: Validation of vascular endothelial growth factor A (VEGF-A) mRNA and transcription factor 20 (TCF20) mRNA in the second cohort of PELS TaqMan gene expression assays of specific primers and MGB probes were purchased from Applied Biosystems for the detection of VEGF-A mRNA (Hs00900054_m1) and TCF20 mRNA (Hs00390028_m1).

Techniques:

Journal: Oncotarget

Article Title: Prognostic significance of Cytokeratin 20-positive lymph node vascular endothelial growth factor A mRNA and chromodomain helicase DNA binding protein 4 in pN0 colorectal cancer patients

doi: 10.18632/oncotarget.23424

Figure Lengend Snippet: ( A ) VEGF-A mRNA and ( B ) TCF20 mRNA copy numbers per 0.05 μg total RNA in CK20+ and CK20- LNs from the 2nd cohort of pN0 CRC patients. The median in each group of subjects was indicated by a black horizontal line.

Article Snippet: Validation of vascular endothelial growth factor A (VEGF-A) mRNA and transcription factor 20 (TCF20) mRNA in the second cohort of PELS TaqMan gene expression assays of specific primers and MGB probes were purchased from Applied Biosystems for the detection of VEGF-A mRNA (Hs00900054_m1) and TCF20 mRNA (Hs00390028_m1).

Techniques:

Journal: International Journal of Oncology

Article Title: Inhibition of androgen-responsive LNCaP prostate cancer cell tumor xenograft growth by dietary phenethyl isothiocyanate correlates with decreased angiogenesis and inhibition of cell attachment

doi: 10.3892/ijo.2012.1335

Figure Lengend Snippet: Western blot analysis of caspase-3 and VEGF in control and PEITC-treated tumors. Western blot analysis of caspase-3 and human VEGF were performed and quantitated as described in Materials and methods. Results were normalized to β-actin levels and expressed as means ± SE (n=4). (A) Caspase-3. (B) VEGF.

Article Snippet: TaqMan gene expression assay primers and probes for human prostate specific antigen (PSA) and glyceraldehydes-3-phosphate dehydrogenase (G3PDH), proliferating cell nuclear antigen (PCNA), Ki-67, integrin β1 (ITGB1), integrin α2 (ITGA2), integrin α5 (ITGA5), integrin α6 (ITGA6), mouse and human vascular endothelial growth factor (VEGF) A, mouse PECAM-1 and mouse and human TATA binding protein were purchased from Applied Biosystems (Foster City, CA).

Techniques: Western Blot

Journal: Scientific Reports

Article Title: Anticancer potentiality of lignan rich fraction of six Flaxseed cultivars

doi: 10.1038/s41598-017-18944-0

Figure Lengend Snippet: Effect of BD,FO, FS and FSM given as diet twice daily for three weeks on VEGF expression in EAC-bearing mice. ( A ) Immunohistochemical staining of VEGF in EAC solid tumor sections (160×). BD (I), FO (II), FS (III) and FSM (IV). ( B ) Quantification of VEGF immunopositive cells to the total area of the microscopic field across six fields. Values were given as means ± SD (n = 6), a,b and c: Significantly different from BD, FO and FS respectively at p

Article Snippet: Assay of vascular endothelial growth factor (VEGF-A) level VEGF was determined in cell culture medium using eBioscience ELISA kit (San Diego, CA, USA).

Techniques: Expressing, Mouse Assay, Immunohistochemistry, Staining

Journal: Scientific Reports

Article Title: Anticancer potentiality of lignan rich fraction of six Flaxseed cultivars

doi: 10.1038/s41598-017-18944-0

Figure Lengend Snippet: Effect of treatment of T47D cells with IC 50 of PFH-G9 (15 μg/ml) or TAM (15 μM) for 48 h on VEGF level. Each point is the mean ± SD of three independent experiments performed in duplicate. a: Significantly different from control group at p

Article Snippet: Assay of vascular endothelial growth factor (VEGF-A) level VEGF was determined in cell culture medium using eBioscience ELISA kit (San Diego, CA, USA).

Techniques:

Journal: Frontiers in Oncology

Article Title: Vascular Endothelial Growth Factor, Irradiation, and Axitinib Have Diverse Effects on Motility and Proliferation of Glioblastoma Multiforme Cells

doi: 10.3389/fonc.2017.00182

Figure Lengend Snippet: Influence of vascular endothelial growth factor (VEGF), axitinib, or irradiation with 2 Gy photons on the motility of glioblastoma multiforme cell lines U-251 and U-373. The motility of the cells was analyzed by time-laps videography. Cells were tracked and analyzed with the ibidi chemotaxis and migration tool. (A) Examples from an image stack of Videography. Some cells migrate fast (2), some slowly (3), some even do not migrate (1). Dividing cells (4) often keep contact over longer periods of time (4a, 4b). Scale bars: 50 µm. (B) Migration of U-373 glioblastoma cells under varied conditions. Depicted are the tracked cells in representative fields of view. The software merges all starting points in the origin to get an explicit view of the paths migrated by the cells. It is clearly visible that the migration is undirected (an advantage of videography over other methods to analyze migration). It is notable that some irradiated cells are able to escape the inhibition by axitinib. (C,D) The motility of U-251 and U-373 cells is increased by VEGF as well as by irradiation. In U-373, a combination of both leads to a significant increase in velocity compared to VEGF alone (D) , in U-251 no additive effects could be observed. In contrast, axitinib diminishes the motility of untreated cells and the elevated motility after irradiation as well. VEGF and axitinib were added in concentrations of 0.1 and 10 µg/ml, respectively. (E,F) 24 h after 2 Gy irradiation the amount of VEGF in the supernatant of U-251 and U-373 was analyzed. In U-251 there were no significant changes detectable, whereas in U-373 cells VEGF was significantly increased (F) . Data are shown as mean ± SEM. Data were tested for significance using one-way ANOVA with Bonferroni multiple comparison post-test. Significant differences are indicated by * p

Article Snippet: Materials VEGF-A (Sigma-Aldrich, V4512, USA) and axitinib (Selleckchem, S1005, USA) was added to cell culture medium in a concentration of 0.1 and 10 µg/ml, respectively.

Techniques: Irradiation, Chemotaxis Assay, Migration, Software, Inhibition

Journal: Frontiers in Oncology

Article Title: Vascular Endothelial Growth Factor, Irradiation, and Axitinib Have Diverse Effects on Motility and Proliferation of Glioblastoma Multiforme Cells

doi: 10.3389/fonc.2017.00182

Figure Lengend Snippet: Expression of vascular endothelial growth factor (VEGF)-R2 ( KDR ) in U-251 and U-373 glioblastoma cell lines. (A,B) VEGF-R1 ( FLT 1) as well as VEGF-R2 ( KDR) are expressed in both cell lines U-251 (A) and U-373 (B) . β -Actin was used as housekeeping gene. PCR products were isolated and confirmed by DNA sequencing. (C,D) Sequence of amplified PCR product for FLT1 (C) and KDR (D) matches their specific database entries ( FLT1 – NM_002019.4; KDR – NM_002253.2) and were proven to be unique in human glioblastoma cell lines U-251 as well as in U-373 by comparison with a database (Blast 2.2, U.S. National Centre for Biotechnology Information, Bethesda, MD, USA) (E) . Semi-quantitative analysis of gene expression normalized to β -Actin and compared to the expression of U-251-FLT (100%). Data are shown as mean ± SEM. n = 3. (F) Both glioblastoma multiforme cell lines express VEGF-R2 (green dots, arrows) in the cytoplasm as well as along the cell membrane. Counterstaining of the actin cytoskeleton is given in red as well as cell nuclei staining with DAPI in blue. Scale bars: 10 µm.

Article Snippet: Materials VEGF-A (Sigma-Aldrich, V4512, USA) and axitinib (Selleckchem, S1005, USA) was added to cell culture medium in a concentration of 0.1 and 10 µg/ml, respectively.

Techniques: Expressing, Polymerase Chain Reaction, Isolation, DNA Sequencing, Sequencing, Amplification, Staining

Journal: Frontiers in Oncology

Article Title: Vascular Endothelial Growth Factor, Irradiation, and Axitinib Have Diverse Effects on Motility and Proliferation of Glioblastoma Multiforme Cells

doi: 10.3389/fonc.2017.00182

Figure Lengend Snippet: Influence of vascular endothelial growth factor (VEGF), axitinib, or irradiation with 2 Gy photons on the proliferation of glioblastoma multiforme cell lines U-251 and U-373. (A,B) Axitinib impairs the proliferation of irradiated and non-irradiated U-251 and U-373 cells. VEGF, irradiation, or the combination of both has no significant effect on cell proliferation. The proliferation of the cells was analyzed by a modified MTS-test. VEGF and axitinib were added in concentrations of 0.1 and 10 µg/ml, respectively. Data are shown as mean ± SEM. Data were tested for significance using one-way ANOVA with Bonferroni multiple comparison post-test. Significant differences are indicated by *** p

Article Snippet: Materials VEGF-A (Sigma-Aldrich, V4512, USA) and axitinib (Selleckchem, S1005, USA) was added to cell culture medium in a concentration of 0.1 and 10 µg/ml, respectively.

Techniques: Irradiation, Modification

Journal: Frontiers in Oncology

Article Title: Vascular Endothelial Growth Factor, Irradiation, and Axitinib Have Diverse Effects on Motility and Proliferation of Glioblastoma Multiforme Cells

doi: 10.3389/fonc.2017.00182

Figure Lengend Snippet: Potential signaling in human glioblastoma cell lines after treatment with vascular endothelial growth factor (VEGF), irradiation and axitinib. VEGF activates multiple pathways including the Cdc42 and PLC pathways concerning cell migration and proliferation. It is supposed that stimulating effects of irradiation are mediated via enhanced synthesis of VEGF. Irradiation elevates VEGF biosynthesis of glioblastoma multiforme cells via MAPK activation. (A) Activation of the Cdc42 pathway by VEGF leads to an increased activation of Arp 2/3, HSP27, and ADF/cofilin resulting in an enhanced motility. Blockade of the VEGF-R2 by axitinib might decrease the activation of the Cdc42 pathway due to theoretical consideration resulting in a crucial decreased cell motility. (B) VEGF activates the PLC pathway which is involved in cell proliferation while axitinib might deactivate this pathway with impairment of cell proliferation. While motility is increased by VEGF, no positive effect on proliferation could be observed. This might be due to fully activated VEGF-pathways even under control conditions. Continuous arrows: evidence of the effect, dotted arrows: assumption of the effect.

Article Snippet: Materials VEGF-A (Sigma-Aldrich, V4512, USA) and axitinib (Selleckchem, S1005, USA) was added to cell culture medium in a concentration of 0.1 and 10 µg/ml, respectively.

Techniques: Irradiation, Planar Chromatography, Migration, Activation Assay

Journal: Scientific Reports

Article Title: Elevated serum interferon γ-induced protein 10 kDa is associated with TAFRO syndrome

doi: 10.1038/srep42316

Figure Lengend Snippet: Steel-Dwass test analysis of between-group differences. ( a ) The median serum IP-10 level was significantly higher in the TAFRO-iMCD group than in the other 2 groups. ( b ) The median serum PDGF-AA level was significantly lower in the TAFRO-iMCD group than in controls and tended to be lower in the TAFRO-iMCD groups than in the iMCD-NOS group. The serum IL-10 ( c ), IL-23 ( d ), and VEGF-A ( e ) levels were significantly higher in the TAFRO-iMCD and iMCD-NOS groups than in the control group. Control: Healthy control, iMCD: iMCD-NOS, TAFRO: TAFRO-iMCD, IP-10: chemokine interferon γ-induced protein 10 kDa, PDGF-AA: platelet-derived growth factor -AA, IL: interleukin, VEGF-A; vascular endothelial growth factor-A, N.S: not significant.

Article Snippet: Measurement of cytokines and chemokines Serum cytokines IL-1β, IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12p70, IL-13, IL-17A, IL-22, IL-23, IL-27, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, platelet-derived growth factor (PDGF)-AA, vascular endothelial growth factor (VEGF)-A, and chemokine interferon γ-induced protein 10 kDa (IP-10) were analyzed using a FlowCytomix™ kit (Bender MedSystems, Vienna, Austria) and MACSQuant® Analyzer (Miltenyi Biotec, Bergisch Gladbach, Germany) in accordance with the manufacturers’ instructions.

Techniques: Derivative Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Matrix Metalloproteinase-2 Impairs Homing of Intracoronary Delivered Mesenchymal Stem Cells in a Porcine Reperfused Myocardial Infarction: Comparison With Intramyocardial Cell Delivery

doi: 10.3389/fbioe.2018.00035

Figure Lengend Snippet: Expression of homing and angiogenic signals of the myocardium 7 days after cardiac transfer of green fluorescent protein (GFP)-Luc-mesenchymal stem cell (MSCs). Fluorescent immunohistochemistry of the bioluminescence positive myocardial areas 7 days after intramyocardial [left panel (A,C,E,G) ] or intracoronary [right panel (B,D,F,H) ] GFP-Luc-MSCs delivery shows increased expression of homing signals cadherin (A,B) , and angiogenic factors fibroblast growth factor 2 (FGF2) (C,D) and vascular endothelial growth factor (VEGF) (E,F) in group IM. Infarct area border zone (G,H) exhibited higher number of myocardial cells and higher level of VEGF expression in group IM (G) . Hoechst staining of the nuclei, 40× magnification. Expression of homing signals cadherin (I) , stromal-derived factor-1alpha (J) , and angiogenic factors FGF2 (K) and VEGF (L) .

Article Snippet: To investigate the homing of the injected GFP-Luc-MSCs, myocardial expression of cadherin (Abcam, Cambridge, UK), and for angiogenic factors, vascular endothelial growth factor (VEGF) (Cusabio, Cologne, Germany) and FGF2 (Lifespan, Seattle, WA, USA) were displayed by immunofluorescence staining.

Techniques: Expressing, Immunohistochemistry, Staining, Derivative Assay