Structured Review

Cell Signaling Technology Inc primary antibodies against vegf
Effect of scutellarin and scutellarin-loaded nanoparticles on the expression of <t>VEGF,</t> <t>VEGFR2</t> and vWF. a Western blot analysis was used to determine the expression of VEGF,VEGFR2 and vWF in the rat retina. Quantitative evaluation of protein expression of VEGF/GAPDH ( b ), VEGFR2/GAPDH ( c ) and vWF/GAPDH ( d ). Data are presented as mean ± SD. *Significantly different from control group ( P
Primary Antibodies Against Vegf, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 91 stars, based on 1 article reviews
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91/100 stars

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1) Product Images from "Enhancement of scutellarin oral delivery efficacy by vitamin B12-modified amphiphilic chitosan derivatives to treat type II diabetes induced-retinopathy"

Article Title: Enhancement of scutellarin oral delivery efficacy by vitamin B12-modified amphiphilic chitosan derivatives to treat type II diabetes induced-retinopathy

Journal: Journal of Nanobiotechnology

doi: 10.1186/s12951-017-0251-z

Effect of scutellarin and scutellarin-loaded nanoparticles on the expression of VEGF, VEGFR2 and vWF. a Western blot analysis was used to determine the expression of VEGF,VEGFR2 and vWF in the rat retina. Quantitative evaluation of protein expression of VEGF/GAPDH ( b ), VEGFR2/GAPDH ( c ) and vWF/GAPDH ( d ). Data are presented as mean ± SD. *Significantly different from control group ( P
Figure Legend Snippet: Effect of scutellarin and scutellarin-loaded nanoparticles on the expression of VEGF, VEGFR2 and vWF. a Western blot analysis was used to determine the expression of VEGF,VEGFR2 and vWF in the rat retina. Quantitative evaluation of protein expression of VEGF/GAPDH ( b ), VEGFR2/GAPDH ( c ) and vWF/GAPDH ( d ). Data are presented as mean ± SD. *Significantly different from control group ( P

Techniques Used: Expressing, Western Blot

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    Cell Signaling Technology Inc primary antibodies against vegf
    Effect of scutellarin and scutellarin-loaded nanoparticles on the expression of <t>VEGF,</t> <t>VEGFR2</t> and vWF. a Western blot analysis was used to determine the expression of VEGF,VEGFR2 and vWF in the rat retina. Quantitative evaluation of protein expression of VEGF/GAPDH ( b ), VEGFR2/GAPDH ( c ) and vWF/GAPDH ( d ). Data are presented as mean ± SD. *Significantly different from control group ( P
    Primary Antibodies Against Vegf, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary antibodies against vegf/product/Cell Signaling Technology Inc
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    primary antibodies against vegf - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

    99
    Cell Signaling Technology Inc antibodies against vegfr 2
    Proposed model for <t>VEGFR-2</t> activation. VEGFR-2 pre-dimerizes in the absence of ligand. Under physiological conditions, corresponding to 10 to 100 VEGFR-2 molcules per square micron, 30 to 60% of the receptors are dimeric. The dimers are stabilized by homotypic contacts in D4-7 in the EC domain, TM interactions that involve amino acids E764, T771, and/or F778, and contacts between the intracellular domains. The EC domain, as a whole, inhibits dimerization. The ligand-free dimers are phosphorylated, to a low degree. ( B ) Ligand binding induces a rotation in the TM helices and the formation of a different TM dimer configuration, such that amino acids E764, T771, and F778 now face the lipid membrane, away from the TM dimer interface. These structural changes increase the separation between the C-termini of the TM helices, promoting an increase in receptor phosphorylation and activation. Contacts between D4 and D7 persist in the ligand-bound state. DOI: http://dx.doi.org/10.7554/eLife.13876.018
    Antibodies Against Vegfr 2, supplied by Cell Signaling Technology Inc, 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/antibodies against vegfr 2/product/Cell Signaling Technology Inc
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    86
    Cell Signaling Technology Inc western blot analyses against vegf
    Naringenin/ β -CD complex significantly inhibits the mRNA expression of key mediators involved in the pathogenesis of CNV in rats. (a)–(f) are real-time PCR analyses of transcript levels of <t>VEGF,</t> <t>COX-2,</t> PI3K, p38MAPK, MMP-2, and MMP-9. GAPDH was used as the invariant control for calculating fold changes in mRNA levels. Group 1: normal rats; group 2: CNV rats without treatment; group 3: CNV rats with naringenin treatment; group 4: CNV rats with naringenin/ β -CD complex treatment. Data are expressed as mean ± SD; # P
    Western Blot Analyses Against 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 antibody against vegf a
    miR-145 attenuated <t>VEGF-A</t> and ANGPT2 in BMECs under OGD conditions ( A ) qRT-PCR analysis was performed to measure relative expression level of VEGF-A in BMECs. ( B ) qRT-PCR analysis was performed to measure relative expression level of ANGPT2 in BMECs. ( C ) Western blot analysis was performed to measure protein levels of VEGF-A and ANGPT2. GAPDH was used as an internal control. BMECs were treated under OGD conditions for 12 h, grouping as OGD(−), OGD(12 h), mimic NC, miR-145 mimic, inhibitor NC, and miR-145 inhibitor. Data represent means ± S.D. based on three independent experiments; * P
    Antibody Against Vegf A, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effect of scutellarin and scutellarin-loaded nanoparticles on the expression of VEGF, VEGFR2 and vWF. a Western blot analysis was used to determine the expression of VEGF,VEGFR2 and vWF in the rat retina. Quantitative evaluation of protein expression of VEGF/GAPDH ( b ), VEGFR2/GAPDH ( c ) and vWF/GAPDH ( d ). Data are presented as mean ± SD. *Significantly different from control group ( P

    Journal: Journal of Nanobiotechnology

    Article Title: Enhancement of scutellarin oral delivery efficacy by vitamin B12-modified amphiphilic chitosan derivatives to treat type II diabetes induced-retinopathy

    doi: 10.1186/s12951-017-0251-z

    Figure Lengend Snippet: Effect of scutellarin and scutellarin-loaded nanoparticles on the expression of VEGF, VEGFR2 and vWF. a Western blot analysis was used to determine the expression of VEGF,VEGFR2 and vWF in the rat retina. Quantitative evaluation of protein expression of VEGF/GAPDH ( b ), VEGFR2/GAPDH ( c ) and vWF/GAPDH ( d ). Data are presented as mean ± SD. *Significantly different from control group ( P

    Article Snippet: Primary antibodies against VEGF, VEGFR2, vWF and Horseradish peroxidase (HRP)-conjugated secondary antibodies were obtained from Cell Signaling Technology (Beverly, MA).

    Techniques: Expressing, Western Blot

    Proposed model for VEGFR-2 activation. VEGFR-2 pre-dimerizes in the absence of ligand. Under physiological conditions, corresponding to 10 to 100 VEGFR-2 molcules per square micron, 30 to 60% of the receptors are dimeric. The dimers are stabilized by homotypic contacts in D4-7 in the EC domain, TM interactions that involve amino acids E764, T771, and/or F778, and contacts between the intracellular domains. The EC domain, as a whole, inhibits dimerization. The ligand-free dimers are phosphorylated, to a low degree. ( B ) Ligand binding induces a rotation in the TM helices and the formation of a different TM dimer configuration, such that amino acids E764, T771, and F778 now face the lipid membrane, away from the TM dimer interface. These structural changes increase the separation between the C-termini of the TM helices, promoting an increase in receptor phosphorylation and activation. Contacts between D4 and D7 persist in the ligand-bound state. DOI: http://dx.doi.org/10.7554/eLife.13876.018

    Journal: eLife

    Article Title: VEGFR-2 conformational switch in response to ligand binding

    doi: 10.7554/eLife.13876

    Figure Lengend Snippet: Proposed model for VEGFR-2 activation. VEGFR-2 pre-dimerizes in the absence of ligand. Under physiological conditions, corresponding to 10 to 100 VEGFR-2 molcules per square micron, 30 to 60% of the receptors are dimeric. The dimers are stabilized by homotypic contacts in D4-7 in the EC domain, TM interactions that involve amino acids E764, T771, and/or F778, and contacts between the intracellular domains. The EC domain, as a whole, inhibits dimerization. The ligand-free dimers are phosphorylated, to a low degree. ( B ) Ligand binding induces a rotation in the TM helices and the formation of a different TM dimer configuration, such that amino acids E764, T771, and F778 now face the lipid membrane, away from the TM dimer interface. These structural changes increase the separation between the C-termini of the TM helices, promoting an increase in receptor phosphorylation and activation. Contacts between D4 and D7 persist in the ligand-bound state. DOI: http://dx.doi.org/10.7554/eLife.13876.018

    Article Snippet: VEGFR-2 total protein levels were assessed using antibodies against VEGFR-2 (55B11; #2479, Cell Signaling Technologies) or the HA tag (3F10, Roche Applied Science).

    Techniques: Activation Assay, Ligand Binding Assay

    Left: Western blots under reducing conditions, for full length C482R VEGFR-2 and for the C482R EC+TM-YFP VEGFR-2 construct. Right: Western blots under non-reducing conditions. No dimeric bands were observed under any conditions. The constitutive dimerization of the C482R mutant is not due to cysteine-induced intermolecular cross-linking mediated by unpaired cysteines. DOI: http://dx.doi.org/10.7554/eLife.13876.017

    Journal: eLife

    Article Title: VEGFR-2 conformational switch in response to ligand binding

    doi: 10.7554/eLife.13876

    Figure Lengend Snippet: Left: Western blots under reducing conditions, for full length C482R VEGFR-2 and for the C482R EC+TM-YFP VEGFR-2 construct. Right: Western blots under non-reducing conditions. No dimeric bands were observed under any conditions. The constitutive dimerization of the C482R mutant is not due to cysteine-induced intermolecular cross-linking mediated by unpaired cysteines. DOI: http://dx.doi.org/10.7554/eLife.13876.017

    Article Snippet: VEGFR-2 total protein levels were assessed using antibodies against VEGFR-2 (55B11; #2479, Cell Signaling Technologies) or the HA tag (3F10, Roche Applied Science).

    Techniques: Western Blot, Construct, Mutagenesis

    Dimerization curves for the wild-type VEGFR-2 in the absence of ligand, the D4→D5 mutant in the absence of ligand, and the D4→D5 mutant in the presence of VEGF-A 121 . DOI: http://dx.doi.org/10.7554/eLife.13876.014

    Journal: eLife

    Article Title: VEGFR-2 conformational switch in response to ligand binding

    doi: 10.7554/eLife.13876

    Figure Lengend Snippet: Dimerization curves for the wild-type VEGFR-2 in the absence of ligand, the D4→D5 mutant in the absence of ligand, and the D4→D5 mutant in the presence of VEGF-A 121 . DOI: http://dx.doi.org/10.7554/eLife.13876.014

    Article Snippet: VEGFR-2 total protein levels were assessed using antibodies against VEGFR-2 (55B11; #2479, Cell Signaling Technologies) or the HA tag (3F10, Roche Applied Science).

    Techniques: Mutagenesis

    Cross-linking of CHO cells expressing full-length wild-type VEGFR-2. Cells were starved for 24 hr to ensure that there was no ligand present. Staining with anti-VEGFR-2 antibodies shows the presence of a glycosylated monomer band at MW ~240 kDa and glycosylated dimer band at MW ~480 kDa. These results support the FRET results that VEGFR-2 forms ligand-independent dimers in the plasma membrane. DOI: http://dx.doi.org/10.7554/eLife.13876.007

    Journal: eLife

    Article Title: VEGFR-2 conformational switch in response to ligand binding

    doi: 10.7554/eLife.13876

    Figure Lengend Snippet: Cross-linking of CHO cells expressing full-length wild-type VEGFR-2. Cells were starved for 24 hr to ensure that there was no ligand present. Staining with anti-VEGFR-2 antibodies shows the presence of a glycosylated monomer band at MW ~240 kDa and glycosylated dimer band at MW ~480 kDa. These results support the FRET results that VEGFR-2 forms ligand-independent dimers in the plasma membrane. DOI: http://dx.doi.org/10.7554/eLife.13876.007

    Article Snippet: VEGFR-2 total protein levels were assessed using antibodies against VEGFR-2 (55B11; #2479, Cell Signaling Technologies) or the HA tag (3F10, Roche Applied Science).

    Techniques: Expressing, Staining

    A conformational change in the TM domain dimer upon ligand binding increases VEGFR-2 phosphorylation. ( A ) FRET efficiency measured as a function of acceptor concentration for EC+TM VEGFR-2, in the absence of ligand and in the presence of 3 μg.ml -1 VEGF-A 121 and VEGF-A 165 a. Two hundred to 500 individual vesicles were imaged in at least three independent experiments. Each data point corresponds to a single vesicle. The stochastic FRET contribution ( King et al., 2014 ) is shown as a solid line. Black stars: FRET data without ligand. ( B ) FRET efficiencies for individual vesicles, corrected for the stochastic FRET contribution. There is no dependence on receptor concentration, indicative of constitutive dimerization. ( C ) Donor concentrations versus acceptor concentrations. ( D ) Intrinsic FRET values, measured for VEGFR-2 EC+TM in the presence of 3 μg.ml -1 of VEGF-A 121 , VEGF-A 165 a, VEGF-A 165 b, VEGF-C, VEGF-E, and VEGF-D. ( E ) Graphical representation of the ligand-induced changes in distance between fluorescent proteins, and the inferred changes in TM domain structures. ( F ) The six VEGF ligands increase VEGFR-2 phosphorylation, to the same extent. A representative Western Blot comparing the phosphorylation of Tyr 1175 in the absence of ligand, and in the presence of six VEGF ligands, at concentrations of 3 μg.ml -1 . Only the top bands, corresponding to mature fully glycosylated receptors, are considered here. Phosphorylation is significantly increased upon ligand addition, with the increase being as high as 10 times. ( G ) Quantification of Western blot results for the fully glycosylated receptors (top bands), in the presence of the six ligands. VEGFR-2 phosphorylation in the presence of the six ligands is very similar. DOI: http://dx.doi.org/10.7554/eLife.13876.009

    Journal: eLife

    Article Title: VEGFR-2 conformational switch in response to ligand binding

    doi: 10.7554/eLife.13876

    Figure Lengend Snippet: A conformational change in the TM domain dimer upon ligand binding increases VEGFR-2 phosphorylation. ( A ) FRET efficiency measured as a function of acceptor concentration for EC+TM VEGFR-2, in the absence of ligand and in the presence of 3 μg.ml -1 VEGF-A 121 and VEGF-A 165 a. Two hundred to 500 individual vesicles were imaged in at least three independent experiments. Each data point corresponds to a single vesicle. The stochastic FRET contribution ( King et al., 2014 ) is shown as a solid line. Black stars: FRET data without ligand. ( B ) FRET efficiencies for individual vesicles, corrected for the stochastic FRET contribution. There is no dependence on receptor concentration, indicative of constitutive dimerization. ( C ) Donor concentrations versus acceptor concentrations. ( D ) Intrinsic FRET values, measured for VEGFR-2 EC+TM in the presence of 3 μg.ml -1 of VEGF-A 121 , VEGF-A 165 a, VEGF-A 165 b, VEGF-C, VEGF-E, and VEGF-D. ( E ) Graphical representation of the ligand-induced changes in distance between fluorescent proteins, and the inferred changes in TM domain structures. ( F ) The six VEGF ligands increase VEGFR-2 phosphorylation, to the same extent. A representative Western Blot comparing the phosphorylation of Tyr 1175 in the absence of ligand, and in the presence of six VEGF ligands, at concentrations of 3 μg.ml -1 . Only the top bands, corresponding to mature fully glycosylated receptors, are considered here. Phosphorylation is significantly increased upon ligand addition, with the increase being as high as 10 times. ( G ) Quantification of Western blot results for the fully glycosylated receptors (top bands), in the presence of the six ligands. VEGFR-2 phosphorylation in the presence of the six ligands is very similar. DOI: http://dx.doi.org/10.7554/eLife.13876.009

    Article Snippet: VEGFR-2 total protein levels were assessed using antibodies against VEGFR-2 (55B11; #2479, Cell Signaling Technologies) or the HA tag (3F10, Roche Applied Science).

    Techniques: Ligand Binding Assay, Concentration Assay, Western Blot

    ( A ) CHO cells do not express VEGFR-2 endogenously. ( B ) CHO cells do not express VEGF endogenously. Here, CHO cells, HEK293T cells, and MECs (microvascular endothelial) cells) were stained for VEGF-A. Lysates were reduced before loading. DOI: http://dx.doi.org/10.7554/eLife.13876.005

    Journal: eLife

    Article Title: VEGFR-2 conformational switch in response to ligand binding

    doi: 10.7554/eLife.13876

    Figure Lengend Snippet: ( A ) CHO cells do not express VEGFR-2 endogenously. ( B ) CHO cells do not express VEGF endogenously. Here, CHO cells, HEK293T cells, and MECs (microvascular endothelial) cells) were stained for VEGF-A. Lysates were reduced before loading. DOI: http://dx.doi.org/10.7554/eLife.13876.005

    Article Snippet: VEGFR-2 total protein levels were assessed using antibodies against VEGFR-2 (55B11; #2479, Cell Signaling Technologies) or the HA tag (3F10, Roche Applied Science).

    Techniques: Staining

    The plasmid constructs used in this study. ( A ) The constructs used in the FRET experiments. The full-length receptors had fluorescent proteins attached to their C-termini via a flexible GGS linker. The truncated receptors had the intracellular domain substituted with a fluorescent protein, which was attached to the TM domain via a longer flexible (GGS) 5 linker. SP: signal peptide, EC: extracellular domain, TM: transmembrane domain, IC: Intracellular domain of VEGFR-2. Fluorescent protein was either YFP or mCherry. Amino acid residue numbers are shown above the constructs. ( B ) The constructs used in the Western blotting experiments. DOI: http://dx.doi.org/10.7554/eLife.13876.003

    Journal: eLife

    Article Title: VEGFR-2 conformational switch in response to ligand binding

    doi: 10.7554/eLife.13876

    Figure Lengend Snippet: The plasmid constructs used in this study. ( A ) The constructs used in the FRET experiments. The full-length receptors had fluorescent proteins attached to their C-termini via a flexible GGS linker. The truncated receptors had the intracellular domain substituted with a fluorescent protein, which was attached to the TM domain via a longer flexible (GGS) 5 linker. SP: signal peptide, EC: extracellular domain, TM: transmembrane domain, IC: Intracellular domain of VEGFR-2. Fluorescent protein was either YFP or mCherry. Amino acid residue numbers are shown above the constructs. ( B ) The constructs used in the Western blotting experiments. DOI: http://dx.doi.org/10.7554/eLife.13876.003

    Article Snippet: VEGFR-2 total protein levels were assessed using antibodies against VEGFR-2 (55B11; #2479, Cell Signaling Technologies) or the HA tag (3F10, Roche Applied Science).

    Techniques: Plasmid Preparation, Construct, Western Blot

    The C482R mutation mimics the effect of bound ligand by promoting a structural change in the EC+TM VEGFR-2 dimer. ( A ) FRET efficiencies determined in individual plasma-membrane-derived vesicles. ( B ) Corrected FRET as a function of receptor concentration. The corrected FRET for the mutant does not depend on receptor concentration, demonstrating that the mutant is a constitutive dimer in the presence and absence of ligand. ( C ) Measured donor versus acceptor concentrations in each vesicle. ( D ) Histograms of measured FRET efficiencies for the mutant, yielding Intrinsic FRET values of ~0.42. ( E ) Western blots showing an increase in phosphorylation due to the C482R mutation, in the absence of ligand, and no further increase upon ligand treatment. The top bands correspond to the mature fully glycosylated form of VEGFR-2. ( F ) Confocal images of VEGF-A 121 -AF594 binding to EC+TM VEGFR-2 C482R in CHO membrane vesicles, demonstrating that the C482R mutant is capable of ligand binding. DOI: http://dx.doi.org/10.7554/eLife.13876.015

    Journal: eLife

    Article Title: VEGFR-2 conformational switch in response to ligand binding

    doi: 10.7554/eLife.13876

    Figure Lengend Snippet: The C482R mutation mimics the effect of bound ligand by promoting a structural change in the EC+TM VEGFR-2 dimer. ( A ) FRET efficiencies determined in individual plasma-membrane-derived vesicles. ( B ) Corrected FRET as a function of receptor concentration. The corrected FRET for the mutant does not depend on receptor concentration, demonstrating that the mutant is a constitutive dimer in the presence and absence of ligand. ( C ) Measured donor versus acceptor concentrations in each vesicle. ( D ) Histograms of measured FRET efficiencies for the mutant, yielding Intrinsic FRET values of ~0.42. ( E ) Western blots showing an increase in phosphorylation due to the C482R mutation, in the absence of ligand, and no further increase upon ligand treatment. The top bands correspond to the mature fully glycosylated form of VEGFR-2. ( F ) Confocal images of VEGF-A 121 -AF594 binding to EC+TM VEGFR-2 C482R in CHO membrane vesicles, demonstrating that the C482R mutant is capable of ligand binding. DOI: http://dx.doi.org/10.7554/eLife.13876.015

    Article Snippet: VEGFR-2 total protein levels were assessed using antibodies against VEGFR-2 (55B11; #2479, Cell Signaling Technologies) or the HA tag (3F10, Roche Applied Science).

    Techniques: Mutagenesis, Derivative Assay, Concentration Assay, Western Blot, Binding Assay, Ligand Binding Assay

    All mutations in the TM domain decrease VEGFR-2 phosphorylation. DOI: http://dx.doi.org/10.7554/eLife.13876.012

    Journal: eLife

    Article Title: VEGFR-2 conformational switch in response to ligand binding

    doi: 10.7554/eLife.13876

    Figure Lengend Snippet: All mutations in the TM domain decrease VEGFR-2 phosphorylation. DOI: http://dx.doi.org/10.7554/eLife.13876.012

    Article Snippet: VEGFR-2 total protein levels were assessed using antibodies against VEGFR-2 (55B11; #2479, Cell Signaling Technologies) or the HA tag (3F10, Roche Applied Science).

    Techniques:

    FRET data, reporting on the effects of the E764I-T771I-F778I and N762I-V769I-G770I sets of mutations, engineered in the EC+TM VEGFR-2 plasmids. DOI: http://dx.doi.org/10.7554/eLife.13876.011

    Journal: eLife

    Article Title: VEGFR-2 conformational switch in response to ligand binding

    doi: 10.7554/eLife.13876

    Figure Lengend Snippet: FRET data, reporting on the effects of the E764I-T771I-F778I and N762I-V769I-G770I sets of mutations, engineered in the EC+TM VEGFR-2 plasmids. DOI: http://dx.doi.org/10.7554/eLife.13876.011

    Article Snippet: VEGFR-2 total protein levels were assessed using antibodies against VEGFR-2 (55B11; #2479, Cell Signaling Technologies) or the HA tag (3F10, Roche Applied Science).

    Techniques:

    Naringenin/ β -CD complex significantly inhibits the mRNA expression of key mediators involved in the pathogenesis of CNV in rats. (a)–(f) are real-time PCR analyses of transcript levels of VEGF, COX-2, PI3K, p38MAPK, MMP-2, and MMP-9. GAPDH was used as the invariant control for calculating fold changes in mRNA levels. Group 1: normal rats; group 2: CNV rats without treatment; group 3: CNV rats with naringenin treatment; group 4: CNV rats with naringenin/ β -CD complex treatment. Data are expressed as mean ± SD; # P

    Journal: BioMed Research International

    Article Title: Preparation of Naringenin/β-Cyclodextrin Complex and Its More Potent Alleviative Effect on Choroidal Neovascularization in Rats

    doi: 10.1155/2014/623509

    Figure Lengend Snippet: Naringenin/ β -CD complex significantly inhibits the mRNA expression of key mediators involved in the pathogenesis of CNV in rats. (a)–(f) are real-time PCR analyses of transcript levels of VEGF, COX-2, PI3K, p38MAPK, MMP-2, and MMP-9. GAPDH was used as the invariant control for calculating fold changes in mRNA levels. Group 1: normal rats; group 2: CNV rats without treatment; group 3: CNV rats with naringenin treatment; group 4: CNV rats with naringenin/ β -CD complex treatment. Data are expressed as mean ± SD; # P

    Article Snippet: The primary antibodies used in western blot analyses against VEGF, COX-2, and p38MAPK were from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Naringenin/ β -CD complex significantly inhibits the protein expression of key mediators involved in the pathogenesis of CNV in rats. (a)–(f) are western blot analyses for protein levels of VEGF, COX-2, PI3K, p38MAPK, MMP-2, and MMP-9 and their quantification. β -Actin was used as an invariant control for equal loading. Group 1: normal rats; group 2: CNV rats without treatment; group 3: CNV rats with naringenin treatment; group 4: CNV rats with naringenin/ β -CD complex treatment. Data are expressed as mean ± SD; # P

    Journal: BioMed Research International

    Article Title: Preparation of Naringenin/β-Cyclodextrin Complex and Its More Potent Alleviative Effect on Choroidal Neovascularization in Rats

    doi: 10.1155/2014/623509

    Figure Lengend Snippet: Naringenin/ β -CD complex significantly inhibits the protein expression of key mediators involved in the pathogenesis of CNV in rats. (a)–(f) are western blot analyses for protein levels of VEGF, COX-2, PI3K, p38MAPK, MMP-2, and MMP-9 and their quantification. β -Actin was used as an invariant control for equal loading. Group 1: normal rats; group 2: CNV rats without treatment; group 3: CNV rats with naringenin treatment; group 4: CNV rats with naringenin/ β -CD complex treatment. Data are expressed as mean ± SD; # P

    Article Snippet: The primary antibodies used in western blot analyses against VEGF, COX-2, and p38MAPK were from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Expressing, Western Blot

    miR-145 attenuated VEGF-A and ANGPT2 in BMECs under OGD conditions ( A ) qRT-PCR analysis was performed to measure relative expression level of VEGF-A in BMECs. ( B ) qRT-PCR analysis was performed to measure relative expression level of ANGPT2 in BMECs. ( C ) Western blot analysis was performed to measure protein levels of VEGF-A and ANGPT2. GAPDH was used as an internal control. BMECs were treated under OGD conditions for 12 h, grouping as OGD(−), OGD(12 h), mimic NC, miR-145 mimic, inhibitor NC, and miR-145 inhibitor. Data represent means ± S.D. based on three independent experiments; * P

    Journal: Bioscience Reports

    Article Title: LncRNA MALAT1 up-regulates VEGF-A and ANGPT2 to promote angiogenesis in brain microvascular endothelial cells against oxygen–glucose deprivation via targetting miR-145

    doi: 10.1042/BSR20180226

    Figure Lengend Snippet: miR-145 attenuated VEGF-A and ANGPT2 in BMECs under OGD conditions ( A ) qRT-PCR analysis was performed to measure relative expression level of VEGF-A in BMECs. ( B ) qRT-PCR analysis was performed to measure relative expression level of ANGPT2 in BMECs. ( C ) Western blot analysis was performed to measure protein levels of VEGF-A and ANGPT2. GAPDH was used as an internal control. BMECs were treated under OGD conditions for 12 h, grouping as OGD(−), OGD(12 h), mimic NC, miR-145 mimic, inhibitor NC, and miR-145 inhibitor. Data represent means ± S.D. based on three independent experiments; * P

    Article Snippet: The membrane was incubated with primary antibody against VEGF-A (Cell Signaling Technology, MA, U.S.A.) and ANGPT2 (Cell Signaling Technology, MA, U.S.A.) for 20 h at 4°C.

    Techniques: Quantitative RT-PCR, Expressing, Western Blot

    LncRNA-MALAT1 up-regulated VEGF-A and ANGPT2 via targetting miR-145 ( A ) qRT-PCR analysis was performed to measure relative expression level of VEGF-A in BMECs. ( B ) qRT-PCR analysis was performed to measure relative expression level of ANGPT2 in BMECs. ( C ) qRT-PCR analysis was used to measure the effects of MALAT1 on the expression level of miR-145 in BMECs. ( D ) Western blot analysis was performed to measure protein levels of VEGF-A and ANGPT2, GAPDH was used as an internal control. ( E ) Conservation of MALAT1 at the binding site of miR-145 was snapshot, 5 nts on 3′-UTR of MALAT1 were replaced in constructed MALAT1 mutant. ( F ) Dual-luciferase reporter assay was used to measure relative luciferase activity. BMECs were treated under OGD conditions for 12 h, grouping as OGD(−), OGD(12 h), si-control, si-MALAT1, pcDNA-control, and pcDNA-MALAT1. Data represent means ± S.D. based on three independent experiments; * P

    Journal: Bioscience Reports

    Article Title: LncRNA MALAT1 up-regulates VEGF-A and ANGPT2 to promote angiogenesis in brain microvascular endothelial cells against oxygen–glucose deprivation via targetting miR-145

    doi: 10.1042/BSR20180226

    Figure Lengend Snippet: LncRNA-MALAT1 up-regulated VEGF-A and ANGPT2 via targetting miR-145 ( A ) qRT-PCR analysis was performed to measure relative expression level of VEGF-A in BMECs. ( B ) qRT-PCR analysis was performed to measure relative expression level of ANGPT2 in BMECs. ( C ) qRT-PCR analysis was used to measure the effects of MALAT1 on the expression level of miR-145 in BMECs. ( D ) Western blot analysis was performed to measure protein levels of VEGF-A and ANGPT2, GAPDH was used as an internal control. ( E ) Conservation of MALAT1 at the binding site of miR-145 was snapshot, 5 nts on 3′-UTR of MALAT1 were replaced in constructed MALAT1 mutant. ( F ) Dual-luciferase reporter assay was used to measure relative luciferase activity. BMECs were treated under OGD conditions for 12 h, grouping as OGD(−), OGD(12 h), si-control, si-MALAT1, pcDNA-control, and pcDNA-MALAT1. Data represent means ± S.D. based on three independent experiments; * P

    Article Snippet: The membrane was incubated with primary antibody against VEGF-A (Cell Signaling Technology, MA, U.S.A.) and ANGPT2 (Cell Signaling Technology, MA, U.S.A.) for 20 h at 4°C.

    Techniques: Quantitative RT-PCR, Expressing, Western Blot, Binding Assay, Construct, Mutagenesis, Luciferase, Reporter Assay, Activity Assay

    miR-145 targetted 3′-UTR of VEGF-A and ANGPT2 ( A ) The luciferase reporter constructs containing the wild-type (WT) or mutant (MUT) 3′-UTR of ANGPT2 or VEGF-A. ( B ) Wild-type VEGF-A or mutant VEGF-A was co-transfected into 293T cells with miR-145 or mimic NC. ( C ) Wild-type ANGPT2 or mutant ANGPT2 was co-transfected into 293T cells with miR-145 or mimic NC. Luciferase activity was determined at 48 h after transfection. Data represent means ± S.D. based on three independent experiments; **P

    Journal: Bioscience Reports

    Article Title: LncRNA MALAT1 up-regulates VEGF-A and ANGPT2 to promote angiogenesis in brain microvascular endothelial cells against oxygen–glucose deprivation via targetting miR-145

    doi: 10.1042/BSR20180226

    Figure Lengend Snippet: miR-145 targetted 3′-UTR of VEGF-A and ANGPT2 ( A ) The luciferase reporter constructs containing the wild-type (WT) or mutant (MUT) 3′-UTR of ANGPT2 or VEGF-A. ( B ) Wild-type VEGF-A or mutant VEGF-A was co-transfected into 293T cells with miR-145 or mimic NC. ( C ) Wild-type ANGPT2 or mutant ANGPT2 was co-transfected into 293T cells with miR-145 or mimic NC. Luciferase activity was determined at 48 h after transfection. Data represent means ± S.D. based on three independent experiments; **P

    Article Snippet: The membrane was incubated with primary antibody against VEGF-A (Cell Signaling Technology, MA, U.S.A.) and ANGPT2 (Cell Signaling Technology, MA, U.S.A.) for 20 h at 4°C.

    Techniques: Luciferase, Construct, Mutagenesis, Transfection, Activity Assay