vegf  (Abcam)

 
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    Name:
    Recombinant VEGF Receptor 3 mouse
    Description:

    Catalog Number:
    ab205902
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    Structured Review

    Abcam vegf
    Synergistic effects of combinations of nicotine (N) with EGF (E), <t>IGF-I</t> (I) or <t>VEGF</t> (V) on proliferation of different types of lung cells. Alive, ie, TBD-negative, cells seeded at a density of 1 × 10 4 per well of a 96-well plate were counted after 24 h of incubation in the absence (intact control) or presence of the optimal doses of nicotine (see text) and 10 ng/ml of each test GF. Some cells were exposed to 20 μM HC-3 ( H ) ± nicotine and some — to nicotine/GF combinations in the presence of αBtx ( B; 1 μM) and/or Mec ( M; 50 μM). All values significantly (p

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    Images

    1) Product Images from "Mechanisms of tumor-promoting activities of nicotine in lung cancer: synergistic effects of cell membrane and mitochondrial nicotinic acetylcholine receptors"

    Article Title: Mechanisms of tumor-promoting activities of nicotine in lung cancer: synergistic effects of cell membrane and mitochondrial nicotinic acetylcholine receptors

    Journal: BMC Cancer

    doi: 10.1186/s12885-015-1158-4

    Synergistic effects of combinations of nicotine (N) with EGF (E), IGF-I (I) or VEGF (V) on proliferation of different types of lung cells. Alive, ie, TBD-negative, cells seeded at a density of 1 × 10 4 per well of a 96-well plate were counted after 24 h of incubation in the absence (intact control) or presence of the optimal doses of nicotine (see text) and 10 ng/ml of each test GF. Some cells were exposed to 20 μM HC-3 ( H ) ± nicotine and some — to nicotine/GF combinations in the presence of αBtx ( B; 1 μM) and/or Mec ( M; 50 μM). All values significantly (p
    Figure Legend Snippet: Synergistic effects of combinations of nicotine (N) with EGF (E), IGF-I (I) or VEGF (V) on proliferation of different types of lung cells. Alive, ie, TBD-negative, cells seeded at a density of 1 × 10 4 per well of a 96-well plate were counted after 24 h of incubation in the absence (intact control) or presence of the optimal doses of nicotine (see text) and 10 ng/ml of each test GF. Some cells were exposed to 20 μM HC-3 ( H ) ± nicotine and some — to nicotine/GF combinations in the presence of αBtx ( B; 1 μM) and/or Mec ( M; 50 μM). All values significantly (p

    Techniques Used: Incubation

    Roles of individual cm-nAChR subtypes in implementing synergy of nicotine with GFs. The SW900 cells transfected with either shRNA-NC (control; taken as 100%) or anti-nAChR shRNA were incubated for 24 h in the absence (1) or presence of combinations of 0.1 μM nicotine with 10 ng/ml of EGF (2) , IGF-I (3) or VEGF (4) , and then subjected to direct counting (TBD-negative cells only). Data are mean + SD from a triplicate sample. Asterisk = p
    Figure Legend Snippet: Roles of individual cm-nAChR subtypes in implementing synergy of nicotine with GFs. The SW900 cells transfected with either shRNA-NC (control; taken as 100%) or anti-nAChR shRNA were incubated for 24 h in the absence (1) or presence of combinations of 0.1 μM nicotine with 10 ng/ml of EGF (2) , IGF-I (3) or VEGF (4) , and then subjected to direct counting (TBD-negative cells only). Data are mean + SD from a triplicate sample. Asterisk = p

    Techniques Used: Transfection, shRNA, Incubation

    2) Product Images from "IQGAP1 Regulates Adult Neural Progenitors In Vivo and Vascular Endothelial Growth Factor-Triggered Neural Progenitor Migration In Vitro"

    Article Title: IQGAP1 Regulates Adult Neural Progenitors In Vivo and Vascular Endothelial Growth Factor-Triggered Neural Progenitor Migration In Vitro

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.0830-07.2007

    IQGAP1 regulates VEGF-dependent neural progenitor cell migration. A , Reverse transcription-PCR (RT) analysis performed on wild-type and Iqgap1 -null mutant neurospheres reveal expression of both VEGF receptors Flk-1 (VEGF-R2) and Flt-1 (VEGF-R1). NP, Neural progenitors. B , Phase-contrast reconstitution of time-lapse imaging of wild-type and Iqgap1 -null mutant-derived neurospheres in response to VEGF (20 ng/ml) stimulation. C , Quantification of the average migration distance ( n = 10) of wild-type (WT) and Iqgap1 -null (KO) neurosphere cells after VEGF stimulation.
    Figure Legend Snippet: IQGAP1 regulates VEGF-dependent neural progenitor cell migration. A , Reverse transcription-PCR (RT) analysis performed on wild-type and Iqgap1 -null mutant neurospheres reveal expression of both VEGF receptors Flk-1 (VEGF-R2) and Flt-1 (VEGF-R1). NP, Neural progenitors. B , Phase-contrast reconstitution of time-lapse imaging of wild-type and Iqgap1 -null mutant-derived neurospheres in response to VEGF (20 ng/ml) stimulation. C , Quantification of the average migration distance ( n = 10) of wild-type (WT) and Iqgap1 -null (KO) neurosphere cells after VEGF stimulation.

    Techniques Used: Migration, Polymerase Chain Reaction, Mutagenesis, Expressing, Imaging, Derivative Assay

    Analysis of IQGAP1 partners in wild-type neurospheres stimulated with VEGF. A , VEGF stimulation enhances colocalization of IQGAP1 with Rac1 at the cell membrane. Unstimulated (control) or VEGF-stimulated neurospheres (VEGF) were double immunostained with anti-IQGAP1 (green) and anti-Rac1 (red) antibodies. Scale bars, 10 μm. B , Characterization of IQGAP1 immunoprecipitates (IP) from neurospheres stimulated with VEGF. Total neurosphere extracts (lane 1) and IQGAP1 immunoprecipitates from neurospheres not stimulated (lane 2) or stimulated with VEGF for 10 min (lanes 3) or 30 min (lanes 4) were analyzed by Western blot with polyclonal IQGAP1 and monoclonal Cdc42, Rac1, and Lis1 antibodies as indicated. Lane 5 is a control (Ctl) lane corresponding to the IQGAP1 immunoprecipitate from iqgap 1-null neurospheres.
    Figure Legend Snippet: Analysis of IQGAP1 partners in wild-type neurospheres stimulated with VEGF. A , VEGF stimulation enhances colocalization of IQGAP1 with Rac1 at the cell membrane. Unstimulated (control) or VEGF-stimulated neurospheres (VEGF) were double immunostained with anti-IQGAP1 (green) and anti-Rac1 (red) antibodies. Scale bars, 10 μm. B , Characterization of IQGAP1 immunoprecipitates (IP) from neurospheres stimulated with VEGF. Total neurosphere extracts (lane 1) and IQGAP1 immunoprecipitates from neurospheres not stimulated (lane 2) or stimulated with VEGF for 10 min (lanes 3) or 30 min (lanes 4) were analyzed by Western blot with polyclonal IQGAP1 and monoclonal Cdc42, Rac1, and Lis1 antibodies as indicated. Lane 5 is a control (Ctl) lane corresponding to the IQGAP1 immunoprecipitate from iqgap 1-null neurospheres.

    Techniques Used: Western Blot, CTL Assay

    3) Product Images from "Overexpression of hypoxia-inducible factor-1α and vascular endothelial growth factor in sacral giant cell tumors and the correlation with tumor microvessel density"

    Article Title: Overexpression of hypoxia-inducible factor-1α and vascular endothelial growth factor in sacral giant cell tumors and the correlation with tumor microvessel density

    Journal: Experimental and Therapeutic Medicine

    doi: 10.3892/etm.2014.1971

    Representative immunohistochemical staining for HIF-1α, VEGF and CD34 expression in (A, C and E) sacral GCT samples, respectively, and (B, D and F) normal sacral tissues, respectively (magnification, ×200; light microscopy). HIF-1α, hypoxia-inducible factor 1α; VEGF, vascular endothelial growth factor; GCT, giant cell tumor.
    Figure Legend Snippet: Representative immunohistochemical staining for HIF-1α, VEGF and CD34 expression in (A, C and E) sacral GCT samples, respectively, and (B, D and F) normal sacral tissues, respectively (magnification, ×200; light microscopy). HIF-1α, hypoxia-inducible factor 1α; VEGF, vascular endothelial growth factor; GCT, giant cell tumor.

    Techniques Used: Immunohistochemistry, Staining, Expressing, Light Microscopy

    Overexpression of HIF-1α and VEGF correlates with an increased MVD value in sacral GCT specimens. Immunohistochemistry scores of (A) HIF-1α and (B) VEGF overexpression in the sacral GCT specimens. (C) Increased MVD values were observed in sacral GCT specimens. Correlations between the relative (D) HIF-1α and (E) VEGF expression levels with the MVD value. P
    Figure Legend Snippet: Overexpression of HIF-1α and VEGF correlates with an increased MVD value in sacral GCT specimens. Immunohistochemistry scores of (A) HIF-1α and (B) VEGF overexpression in the sacral GCT specimens. (C) Increased MVD values were observed in sacral GCT specimens. Correlations between the relative (D) HIF-1α and (E) VEGF expression levels with the MVD value. P

    Techniques Used: Over Expression, Immunohistochemistry, Expressing

    Representative western blot analysis for HIF-1α, VEGF and CD34 expression in sacral GCT specimens. (A) HIF-1α, VEGF and CD34 protein expression levels were determined in sacral GCT and normal samples (n=22) using western blot analysis. Relative expression levels of (B) HIF-1α, (C) VEGF and (D) CD34 against GAPDH in the sacral GCT samples. * P
    Figure Legend Snippet: Representative western blot analysis for HIF-1α, VEGF and CD34 expression in sacral GCT specimens. (A) HIF-1α, VEGF and CD34 protein expression levels were determined in sacral GCT and normal samples (n=22) using western blot analysis. Relative expression levels of (B) HIF-1α, (C) VEGF and (D) CD34 against GAPDH in the sacral GCT samples. * P

    Techniques Used: Western Blot, Expressing

    4) Product Images from "Administered circulating microparticles derived from lung cancer patients markedly improved angiogenesis, blood flow and ischemic recovery in rat critical limb ischemia"

    Article Title: Administered circulating microparticles derived from lung cancer patients markedly improved angiogenesis, blood flow and ischemic recovery in rat critical limb ischemia

    Journal: Journal of Translational Medicine

    doi: 10.1186/s12967-015-0381-8

    Proposed mechanisms underlying the positive therapeutic effects of circulating microparticles on critical limb ischemia. Lc-MPs = lung cancer-derived circulating microparticles; NO = nitric oxide; VEGFR2 = vascular endothelial growth factor receptor 2; SDF-1α = stromal cell-derived factor 1 alpha; ANGPT = angiopoietin; eNOS = endothelial nitric oxide synthase; HGF = hepatocyte growth factor; vWF = von Willebrand factor; IFBF = Ratio of ischemic to normal blood flow; PARP = poly (ADP-ribose) polymerase; TGF-β = transforming growth factor beta; BMP = bone morphogenesis protein.
    Figure Legend Snippet: Proposed mechanisms underlying the positive therapeutic effects of circulating microparticles on critical limb ischemia. Lc-MPs = lung cancer-derived circulating microparticles; NO = nitric oxide; VEGFR2 = vascular endothelial growth factor receptor 2; SDF-1α = stromal cell-derived factor 1 alpha; ANGPT = angiopoietin; eNOS = endothelial nitric oxide synthase; HGF = hepatocyte growth factor; vWF = von Willebrand factor; IFBF = Ratio of ischemic to normal blood flow; PARP = poly (ADP-ribose) polymerase; TGF-β = transforming growth factor beta; BMP = bone morphogenesis protein.

    Techniques Used: Derivative Assay, Flow Cytometry

    5) Product Images from "Cellular and Matrix Response of the Mandibular Condylar Cartilage to Botulinum Toxin"

    Article Title: Cellular and Matrix Response of the Mandibular Condylar Cartilage to Botulinum Toxin

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0164599

    Decreased expression of SMAD1/5/8 and VEFG in the Botox injected side MCC and subchondral bone. Immunohistochemistry for SMAD1/5/8 in sagittal sections of control (A) and Botox (B) injected side condyles. VEGF Immunohistochemistry in sections of control (C) and Botox (D) injected side condyles. Scale bar = 500μm.
    Figure Legend Snippet: Decreased expression of SMAD1/5/8 and VEFG in the Botox injected side MCC and subchondral bone. Immunohistochemistry for SMAD1/5/8 in sagittal sections of control (A) and Botox (B) injected side condyles. VEGF Immunohistochemistry in sections of control (C) and Botox (D) injected side condyles. Scale bar = 500μm.

    Techniques Used: Expressing, Injection, Immunohistochemistry

    6) Product Images from "Dramatic enhancement of the detection limits of bioassays via ultrafast deposition of polydopamine"

    Article Title: Dramatic enhancement of the detection limits of bioassays via ultrafast deposition of polydopamine

    Journal: Nature biomedical engineering

    doi: 10.1038/s41551-017-0082

    ELISA and lateral flow strips with EASE ( a ) Schematic illustration of the signal enhancement process. Upon target detection, a layer of PDA is coated around the target molecule, which allows a large number of HRP to adsorb. These HRP molecules, in turn, catalyze the substrate (TMB) conversion at a significantly enhanced rate. ( b ) Visual assessment of the detection sensitivity of ELISA-EASE using mouse IgG as a model target in comparison with conventional ELISA. Colored solutions are visualized in ELISA-EASE wells at target concentrations as low as 10 −13 g ml −1 , while the conventional assay only produces detectable colors at 10 −8 to 10 −9 g ml −1 concentration range. ( c ) Quantitative comparison using values obtained from a plate reader shows the assay response over the full target concentration range (left) and a zoomed-in range close to the essays’ LODs (right). Improvement of approximately 3 orders of magnitude is seen. Error bars, s.d. over three replicates. ( d ) ELISA working curves for four targets, HIV p24, KLK3, CRP, VEGF, with or without EASE. Error bars, s.d. over three replicates. ( e ) The average of LOD improvements for all four targets is around 1,200 fold. ( f ) Strip tests of HIV p24 with or without EASE. Positive signals (indicated by the red arrow) are observed at 10 ng ml −1 and 10 pg ml −1 for the EASE-strips, but the conventional strips can only detect down to 10 ng ml −1 . Each strip represents three replicates.
    Figure Legend Snippet: ELISA and lateral flow strips with EASE ( a ) Schematic illustration of the signal enhancement process. Upon target detection, a layer of PDA is coated around the target molecule, which allows a large number of HRP to adsorb. These HRP molecules, in turn, catalyze the substrate (TMB) conversion at a significantly enhanced rate. ( b ) Visual assessment of the detection sensitivity of ELISA-EASE using mouse IgG as a model target in comparison with conventional ELISA. Colored solutions are visualized in ELISA-EASE wells at target concentrations as low as 10 −13 g ml −1 , while the conventional assay only produces detectable colors at 10 −8 to 10 −9 g ml −1 concentration range. ( c ) Quantitative comparison using values obtained from a plate reader shows the assay response over the full target concentration range (left) and a zoomed-in range close to the essays’ LODs (right). Improvement of approximately 3 orders of magnitude is seen. Error bars, s.d. over three replicates. ( d ) ELISA working curves for four targets, HIV p24, KLK3, CRP, VEGF, with or without EASE. Error bars, s.d. over three replicates. ( e ) The average of LOD improvements for all four targets is around 1,200 fold. ( f ) Strip tests of HIV p24 with or without EASE. Positive signals (indicated by the red arrow) are observed at 10 ng ml −1 and 10 pg ml −1 for the EASE-strips, but the conventional strips can only detect down to 10 ng ml −1 . Each strip represents three replicates.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Flow Cytometry, Concentration Assay, Stripping Membranes

    7) Product Images from "Endothelial progenitor cells improve the therapeutic effect of mesenchymal stem cell sheets on irradiated bone defect repair in a rat model"

    Article Title: Endothelial progenitor cells improve the therapeutic effect of mesenchymal stem cell sheets on irradiated bone defect repair in a rat model

    Journal: Journal of Translational Medicine

    doi: 10.1186/s12967-018-1517-4

    Subcutaneous osteogenesis of cell sheets. Representative H E (×200) and immunohistochemical staining images (×200) of BMP-2, OCN, VEGF and CD31. New bone formation (NB), osteoid (OS), blood vessels (asterisk) are indicated in the images. Black arrows indicate positive staining cells, red arrows indicate positive staining bone matrix
    Figure Legend Snippet: Subcutaneous osteogenesis of cell sheets. Representative H E (×200) and immunohistochemical staining images (×200) of BMP-2, OCN, VEGF and CD31. New bone formation (NB), osteoid (OS), blood vessels (asterisk) are indicated in the images. Black arrows indicate positive staining cells, red arrows indicate positive staining bone matrix

    Techniques Used: Immunohistochemistry, Staining

    Osteogenic genes and proteins expression of cell sheets after osteogenic differentiation. a Gene expression of Runx2, Alp, Bmp - 2, Ocn and Vegf after 0, 3 and 7 days of osteogenic induction. b Western blot analysis of RUNX2, ALP, BMP-2, OCN, VEGF and GAPDH after 7 days of osteogenic induction and the quantitative analysis of the protein bands. Data are shown as mean ± SD and analyzed by Student t-test, n = 3; *p
    Figure Legend Snippet: Osteogenic genes and proteins expression of cell sheets after osteogenic differentiation. a Gene expression of Runx2, Alp, Bmp - 2, Ocn and Vegf after 0, 3 and 7 days of osteogenic induction. b Western blot analysis of RUNX2, ALP, BMP-2, OCN, VEGF and GAPDH after 7 days of osteogenic induction and the quantitative analysis of the protein bands. Data are shown as mean ± SD and analyzed by Student t-test, n = 3; *p

    Techniques Used: Expressing, ALP Assay, Western Blot

    8) Product Images from "Expression profile of cathepsins indicates the potential of cathepsins B and D as prognostic factors in breast cancer patients"

    Article Title: Expression profile of cathepsins indicates the potential of cathepsins B and D as prognostic factors in breast cancer patients

    Journal: Oncology Letters

    doi: 10.3892/ol.2015.3960

    Detection of cathepsins B, D, G, K, L and V as well as RECK and VEGF proteins in breast cancer tissue specimens using immunohistochemistry. Representative images were selected to reflect the expression of (A and B) cathepsin B, (C and D) cathepsin C, (E and F) cathepsin D, (G and H) cathepsin K, (I and J) cathepsin L, (K and L) RECK, (M and N) cathepsin V expression and (O and P) VEGF in breast cancer tissues. Magnification, ×200. RECK, reversion-inducing cysteine-rich protein with Kazal motifs; VEGF, vascular endothelial growth factor.
    Figure Legend Snippet: Detection of cathepsins B, D, G, K, L and V as well as RECK and VEGF proteins in breast cancer tissue specimens using immunohistochemistry. Representative images were selected to reflect the expression of (A and B) cathepsin B, (C and D) cathepsin C, (E and F) cathepsin D, (G and H) cathepsin K, (I and J) cathepsin L, (K and L) RECK, (M and N) cathepsin V expression and (O and P) VEGF in breast cancer tissues. Magnification, ×200. RECK, reversion-inducing cysteine-rich protein with Kazal motifs; VEGF, vascular endothelial growth factor.

    Techniques Used: Immunohistochemistry, Expressing

    9) Product Images from "IL13Rα2 siRNA inhibited cell proliferation, induced cell apoptosis, and suppressed cell invasion in papillary thyroid carcinoma cells"

    Article Title: IL13Rα2 siRNA inhibited cell proliferation, induced cell apoptosis, and suppressed cell invasion in papillary thyroid carcinoma cells

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S153703

    Higher expression level of IL13Rα2 and VEGF were detected in PTC tissues. Notes: Higher mRNA levels of IL13Rα2 ( A ) and VEGF ( B ) were measured in PTC tissue samples (n=45) by using real-time PCR compared with paracancerous tissues (n=45). ** P
    Figure Legend Snippet: Higher expression level of IL13Rα2 and VEGF were detected in PTC tissues. Notes: Higher mRNA levels of IL13Rα2 ( A ) and VEGF ( B ) were measured in PTC tissue samples (n=45) by using real-time PCR compared with paracancerous tissues (n=45). ** P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    IL13Rα2 siRNA inhibited cell invasion in TPC-1 cells. Notes: ( A ) Downregulation of invasion rate was detected in TPC-1 and ARO cells after the transfection with IL13Rα2 siRNAs (siRNA-1 and siRNA-2). ( B ) Expression levels of VEGF, VEGFR2, MMP2, and MMP9 in TPC-1 and ARO cells were measured by using Western blot analysis. ** P
    Figure Legend Snippet: IL13Rα2 siRNA inhibited cell invasion in TPC-1 cells. Notes: ( A ) Downregulation of invasion rate was detected in TPC-1 and ARO cells after the transfection with IL13Rα2 siRNAs (siRNA-1 and siRNA-2). ( B ) Expression levels of VEGF, VEGFR2, MMP2, and MMP9 in TPC-1 and ARO cells were measured by using Western blot analysis. ** P

    Techniques Used: Transfection, Expressing, Western Blot

    10) Product Images from "VEGF promotes gastric cancer development by upregulating CRMP4"

    Article Title: VEGF promotes gastric cancer development by upregulating CRMP4

    Journal: Oncotarget

    doi: 10.18632/oncotarget.7717

    Immunohistochemical analysis of the expression levels of VEGF, VEGFR2 and CRMP4 in tumor sections from gastric cancer patients A. Immunohistochemical detection of VEGF, VEGFR2, and CRMP4 expression levels in tumor and tumor-adjacent tissues collected from gastric cancer patients. B. The relative intensities of VEGF, VEGFR2 and CRMP4 expression in 10 pairs of tumor and tumor-adjacent tissues were evaluated by IPP based on the IHC results. The results analyzed with ANOVA are expressed as means ± SD. The protein expression levels of VEGF and CRMP4 were significantly increased in gastric cancer tissues compared with tumor-adjacent tissues, whereas the VEGFR2 expression levels did not display any significant increase.
    Figure Legend Snippet: Immunohistochemical analysis of the expression levels of VEGF, VEGFR2 and CRMP4 in tumor sections from gastric cancer patients A. Immunohistochemical detection of VEGF, VEGFR2, and CRMP4 expression levels in tumor and tumor-adjacent tissues collected from gastric cancer patients. B. The relative intensities of VEGF, VEGFR2 and CRMP4 expression in 10 pairs of tumor and tumor-adjacent tissues were evaluated by IPP based on the IHC results. The results analyzed with ANOVA are expressed as means ± SD. The protein expression levels of VEGF and CRMP4 were significantly increased in gastric cancer tissues compared with tumor-adjacent tissues, whereas the VEGFR2 expression levels did not display any significant increase.

    Techniques Used: Immunohistochemistry, Expressing

    Expression of CRMP4, VEGFR2 and VEGF in various gastric carcinoma cell lines A. The expression level of VEGF was quantified by ELISA in six gastric carcinoma cell lines. B. Western blot analysis of CRMP4 and VEGFR2 protein expression in each cell line. GADPH was used as a loading control. The expression levels of CRMP4 and VEGFR2 were significantly increased in HGC27 and SGC7901 compared with the other gastric carcinoma cell lines.
    Figure Legend Snippet: Expression of CRMP4, VEGFR2 and VEGF in various gastric carcinoma cell lines A. The expression level of VEGF was quantified by ELISA in six gastric carcinoma cell lines. B. Western blot analysis of CRMP4 and VEGFR2 protein expression in each cell line. GADPH was used as a loading control. The expression levels of CRMP4 and VEGFR2 were significantly increased in HGC27 and SGC7901 compared with the other gastric carcinoma cell lines.

    Techniques Used: Expressing, Enzyme-linked Immunosorbent Assay, Western Blot

    Quantification of VEGF, VEGFR2 and CRMP4 proteins and mRNA levels in gastric cancer tissues A. Western blot analysis showing increased expression levels of CRMP4, VEGF and VEGF2. B. Relative expression of CRMP4 and VEGF by western blot analysis. C. qRT-PCR to detect the relative mRNA expression levels of CRMP4, VEGF and VEGF2 in gastric cancer and tumor-adjacent tissues. All experiments were performed in triplicate. The results from 3 pairs of specimens analyzed by ANOVA are expressed as means ± SD. * P
    Figure Legend Snippet: Quantification of VEGF, VEGFR2 and CRMP4 proteins and mRNA levels in gastric cancer tissues A. Western blot analysis showing increased expression levels of CRMP4, VEGF and VEGF2. B. Relative expression of CRMP4 and VEGF by western blot analysis. C. qRT-PCR to detect the relative mRNA expression levels of CRMP4, VEGF and VEGF2 in gastric cancer and tumor-adjacent tissues. All experiments were performed in triplicate. The results from 3 pairs of specimens analyzed by ANOVA are expressed as means ± SD. * P

    Techniques Used: Western Blot, Expressing, Quantitative RT-PCR

    11) Product Images from "Cyclic stretch induced-retinal pigment epithelial cell apoptosis and cytokine changes"

    Article Title: Cyclic stretch induced-retinal pigment epithelial cell apoptosis and cytokine changes

    Journal: BMC Ophthalmology

    doi: 10.1186/s12886-017-0606-0

    Western blotting analysis of IL6 and VEGF expression. a Colored bands of IL6 and VEGF; b Expression levels of IL6 and VEGF. The expression levels of both IL6 and VEGF increased with the duration of the stretch, reaching their peak levels at 20 h but showed significant reduction at 24 h ( p
    Figure Legend Snippet: Western blotting analysis of IL6 and VEGF expression. a Colored bands of IL6 and VEGF; b Expression levels of IL6 and VEGF. The expression levels of both IL6 and VEGF increased with the duration of the stretch, reaching their peak levels at 20 h but showed significant reduction at 24 h ( p

    Techniques Used: Western Blot, Expressing

    12) Product Images from "P311 Deficiency Leads to Attenuated Angiogenesis in Cutaneous Wound Healing"

    Article Title: P311 Deficiency Leads to Attenuated Angiogenesis in Cutaneous Wound Healing

    Journal: Frontiers in Physiology

    doi: 10.3389/fphys.2017.01004

    Effect of P311 expression on wound angiogenesis. (A) IHC staining of granulation tissues (day 5) for CD31, VEGF, and TGFβ1. (B) Western blot showed the expression of CD31, VEGF, and TGFβ1 in each group. Quantitation of the protein levels of CD31, VEGF, and TGFβ1 in each group (day 5). Levels of VEGF (C) and TGFβ1 (D) releasing during skin wound (day 5) were measured by sandwich ELISA. Data were expressed as means ± SD . n = 5 per group. * P
    Figure Legend Snippet: Effect of P311 expression on wound angiogenesis. (A) IHC staining of granulation tissues (day 5) for CD31, VEGF, and TGFβ1. (B) Western blot showed the expression of CD31, VEGF, and TGFβ1 in each group. Quantitation of the protein levels of CD31, VEGF, and TGFβ1 in each group (day 5). Levels of VEGF (C) and TGFβ1 (D) releasing during skin wound (day 5) were measured by sandwich ELISA. Data were expressed as means ± SD . n = 5 per group. * P

    Techniques Used: Expressing, Immunohistochemistry, Staining, Western Blot, Quantitation Assay, Sandwich ELISA

    13) Product Images from "Angiogenesis for tumor vascular normalization of Endostar on hepatoma 22 tumor-bearing mice is involved in the immune response"

    Article Title: Angiogenesis for tumor vascular normalization of Endostar on hepatoma 22 tumor-bearing mice is involved in the immune response

    Journal: Oncology Letters

    doi: 10.3892/ol.2018.7734

    Effects of Endostar on the mRNA levels of (A) HIF-1α and (B) VEGF in liver tissue of hepatoma 22-bearing C57BL/6 mice. All experiments were performed at least three times. Results are presented as the mean ± standard deviation (n=6). # P
    Figure Legend Snippet: Effects of Endostar on the mRNA levels of (A) HIF-1α and (B) VEGF in liver tissue of hepatoma 22-bearing C57BL/6 mice. All experiments were performed at least three times. Results are presented as the mean ± standard deviation (n=6). # P

    Techniques Used: Mouse Assay, Standard Deviation

    (A) Effects of Endostar on the expression of angiogenesis-associated proteins in mouse liver tissues during treatment. Protein from liver tissues was extracted and protein expression of (B) VEGF, (C) CD31, (D) HIF-1α, (E) MMP-2 and (F) MMP-9 was analyzed using Image-Pro Plus 6.0. All experiments were performed at least three times. Results are presented as the mean ± standard deviation (n=6). # P
    Figure Legend Snippet: (A) Effects of Endostar on the expression of angiogenesis-associated proteins in mouse liver tissues during treatment. Protein from liver tissues was extracted and protein expression of (B) VEGF, (C) CD31, (D) HIF-1α, (E) MMP-2 and (F) MMP-9 was analyzed using Image-Pro Plus 6.0. All experiments were performed at least three times. Results are presented as the mean ± standard deviation (n=6). # P

    Techniques Used: Expressing, Standard Deviation

    14) Product Images from "High Intensity Interval and Endurance Training Have Opposing Effects on Markers of Heart Failure and Cardiac Remodeling in Hypertensive Rats"

    Article Title: High Intensity Interval and Endurance Training Have Opposing Effects on Markers of Heart Failure and Cardiac Remodeling in Hypertensive Rats

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0121138

    Western blot analysis of eNOS, HIF1α, and VEGF. A. Representative blots; α-tubulin is presented as a loading control. B. Density quantifications of eNOS protein in LV of low sodium sedentary (LS-SED), high sodium SED (HS-SED), HS endurance training (HS-ET) and HS high intensity interval training (HS-HIIT), demonstrating a significant increase as a result of ET, * vs. all other groups; P
    Figure Legend Snippet: Western blot analysis of eNOS, HIF1α, and VEGF. A. Representative blots; α-tubulin is presented as a loading control. B. Density quantifications of eNOS protein in LV of low sodium sedentary (LS-SED), high sodium SED (HS-SED), HS endurance training (HS-ET) and HS high intensity interval training (HS-HIIT), demonstrating a significant increase as a result of ET, * vs. all other groups; P

    Techniques Used: Western Blot

    15) Product Images from "FM19G11 inhibits O6‐methylguanine DNA‐methyltransferase expression under both hypoxic and normoxic conditions. FM19G11 inhibits O6‐methylguanine DNA‐methyltransferase expression under both hypoxic and normoxic conditions"

    Article Title: FM19G11 inhibits O6‐methylguanine DNA‐methyltransferase expression under both hypoxic and normoxic conditions. FM19G11 inhibits O6‐methylguanine DNA‐methyltransferase expression under both hypoxic and normoxic conditions

    Journal: Cancer Medicine

    doi: 10.1002/cam4.1551

    Under hypoxic conditions, HIF ‐1α was expressed in both GBM ‐ XD and T98G cells. After FM 19G11 treatment, the levels of HIF ‐1α and its target genes EPO and VEGF were decreased, and the change was similar to that of MGMT
    Figure Legend Snippet: Under hypoxic conditions, HIF ‐1α was expressed in both GBM ‐ XD and T98G cells. After FM 19G11 treatment, the levels of HIF ‐1α and its target genes EPO and VEGF were decreased, and the change was similar to that of MGMT

    Techniques Used:

    16) Product Images from "FOXP3 inhibits angiogenesis by downregulating VEGF in breast cancer"

    Article Title: FOXP3 inhibits angiogenesis by downregulating VEGF in breast cancer

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-018-0790-8

    VEGF is involved in the FOXP3-mediated inhibition of angiogenesis. a The tube formation activity of HUVECs that were treated with control medium, the culture supernatant of MAD-MB-231 cells (control supernatant), the culture supernatant of FOXP3-overexpressing MAD-MB-231 cells (FOXP3 supernatant), or the culture supernatant of FOXP3-overexpressing MAD-MB-231 cells supplemented with VEGF (FOXP3 supernatant + VEGF). Scale bar, 50 μm. b Quantitation of HUVEC tubulogenesis in the different groups in ( a ). HUVEC spheroid sprouting activity was determined by 3D culture. c Representative images of spheroid sprouting. Scale bar, 100 μm. d Quantitation of the cumulative sprout length (CSL) in different groups. b , d ANOVA with Tukey’s post hoc test
    Figure Legend Snippet: VEGF is involved in the FOXP3-mediated inhibition of angiogenesis. a The tube formation activity of HUVECs that were treated with control medium, the culture supernatant of MAD-MB-231 cells (control supernatant), the culture supernatant of FOXP3-overexpressing MAD-MB-231 cells (FOXP3 supernatant), or the culture supernatant of FOXP3-overexpressing MAD-MB-231 cells supplemented with VEGF (FOXP3 supernatant + VEGF). Scale bar, 50 μm. b Quantitation of HUVEC tubulogenesis in the different groups in ( a ). HUVEC spheroid sprouting activity was determined by 3D culture. c Representative images of spheroid sprouting. Scale bar, 100 μm. d Quantitation of the cumulative sprout length (CSL) in different groups. b , d ANOVA with Tukey’s post hoc test

    Techniques Used: Inhibition, Activity Assay, Quantitation Assay

    Inverse correlation between FOXP3 and VEGF expression in human breast cancer samples. a Representative immunohistochemical images of nuclear FOXP3 and VEGF expression in breast cancer specimens. Scale bar, 100 μm (×10) and 20 μm (×40). b A significant negative correlation between nuclear FOXP3 and VEGF expression was found in the breast cancer specimens. b Spearman test
    Figure Legend Snippet: Inverse correlation between FOXP3 and VEGF expression in human breast cancer samples. a Representative immunohistochemical images of nuclear FOXP3 and VEGF expression in breast cancer specimens. Scale bar, 100 μm (×10) and 20 μm (×40). b A significant negative correlation between nuclear FOXP3 and VEGF expression was found in the breast cancer specimens. b Spearman test

    Techniques Used: Expressing, Immunohistochemistry

    FOXP3 downregulates VEGF expression in breast cancer. Real-time PCR was performed to detect the transcription levels of VEGF in breast cancer cell lines with the gain or loss of FOXP3 expression. Transfection of pcDNA3.1-FOXP3 into ( a ) MCF-7, ( b ) T47D or ( c ) MAD-MB-231 cells. Transfection of FOXP3 shRNA into ( d ) MCF-7 or ( e ) T47D cells. Western blotting was performed to detect the expression of VEGF in breast cancer cell lines following the gain or loss of FOXP3 expression. Transfection of pcDNA3.1-FOXP3 into ( f ) MCF-7, ( g ) T47D or ( h ) MAD-MB-231 cells. Transfection of FOXP3 shRNA into ( i ) MCF-7 or ( j ) T47D cells. k Evaluation of FOXP3 and VEGF expression in MAD-MB-231 cells by confocal microscopy (control or FOXP3 overexpression). Scale bar, 20 μm. l Orthotopic injection of MDA-MB-231 cells was performed to generate xenografts, and adenoviruses carrying FOXP3 or control cDNA were injected into the tumors when their volume reached ~50 mm 3 ( n = 5). Serum VEGF levels of mice bearing control or FOXP3-overexpressing tumors were detected by ELISA. a – l Student’s t test
    Figure Legend Snippet: FOXP3 downregulates VEGF expression in breast cancer. Real-time PCR was performed to detect the transcription levels of VEGF in breast cancer cell lines with the gain or loss of FOXP3 expression. Transfection of pcDNA3.1-FOXP3 into ( a ) MCF-7, ( b ) T47D or ( c ) MAD-MB-231 cells. Transfection of FOXP3 shRNA into ( d ) MCF-7 or ( e ) T47D cells. Western blotting was performed to detect the expression of VEGF in breast cancer cell lines following the gain or loss of FOXP3 expression. Transfection of pcDNA3.1-FOXP3 into ( f ) MCF-7, ( g ) T47D or ( h ) MAD-MB-231 cells. Transfection of FOXP3 shRNA into ( i ) MCF-7 or ( j ) T47D cells. k Evaluation of FOXP3 and VEGF expression in MAD-MB-231 cells by confocal microscopy (control or FOXP3 overexpression). Scale bar, 20 μm. l Orthotopic injection of MDA-MB-231 cells was performed to generate xenografts, and adenoviruses carrying FOXP3 or control cDNA were injected into the tumors when their volume reached ~50 mm 3 ( n = 5). Serum VEGF levels of mice bearing control or FOXP3-overexpressing tumors were detected by ELISA. a – l Student’s t test

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Transfection, shRNA, Western Blot, Confocal Microscopy, Over Expression, Injection, Multiple Displacement Amplification, Mouse Assay, Enzyme-linked Immunosorbent Assay

    FOXP3 is a transcriptional suppressor of VEGF. a FOXP3 suppresses VEGF promoter activity, as evaluated by dual-luciferase reporter assays. b The top panel depicts schematic diagrams of the regions amplified by the ChIP primers. The bottom panel shows the amount of DNA precipitated by either the anti-FOXP3 antibody or control IgG; the results are expressed as a percentage of the input genomic DNA from MDA-MB-231 cells. c FOXP3-mediated suppression of the VEGF promoter requires forkhead-binding motifs, as evaluated by dual-luciferase reporter assays. Relative truncation of the VEGF promoter is illustrated in the left panel. Luciferase activity was detected in cells transfected with the truncated VEGF promoter in the right panel. a , c ANOVA with Tukey’s post hoc test
    Figure Legend Snippet: FOXP3 is a transcriptional suppressor of VEGF. a FOXP3 suppresses VEGF promoter activity, as evaluated by dual-luciferase reporter assays. b The top panel depicts schematic diagrams of the regions amplified by the ChIP primers. The bottom panel shows the amount of DNA precipitated by either the anti-FOXP3 antibody or control IgG; the results are expressed as a percentage of the input genomic DNA from MDA-MB-231 cells. c FOXP3-mediated suppression of the VEGF promoter requires forkhead-binding motifs, as evaluated by dual-luciferase reporter assays. Relative truncation of the VEGF promoter is illustrated in the left panel. Luciferase activity was detected in cells transfected with the truncated VEGF promoter in the right panel. a , c ANOVA with Tukey’s post hoc test

    Techniques Used: Activity Assay, Luciferase, Amplification, Chromatin Immunoprecipitation, Multiple Displacement Amplification, Binding Assay, Transfection

    17) Product Images from "FOXP3 inhibits angiogenesis by downregulating VEGF in breast cancer"

    Article Title: FOXP3 inhibits angiogenesis by downregulating VEGF in breast cancer

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-018-0790-8

    VEGF is involved in the FOXP3-mediated inhibition of angiogenesis. a The tube formation activity of HUVECs that were treated with control medium, the culture supernatant of MAD-MB-231 cells (control supernatant), the culture supernatant of FOXP3-overexpressing MAD-MB-231 cells (FOXP3 supernatant), or the culture supernatant of FOXP3-overexpressing MAD-MB-231 cells supplemented with VEGF (FOXP3 supernatant + VEGF). Scale bar, 50 μm. b Quantitation of HUVEC tubulogenesis in the different groups in ( a ). HUVEC spheroid sprouting activity was determined by 3D culture. c Representative images of spheroid sprouting. Scale bar, 100 μm. d Quantitation of the cumulative sprout length (CSL) in different groups. b , d ANOVA with Tukey’s post hoc test
    Figure Legend Snippet: VEGF is involved in the FOXP3-mediated inhibition of angiogenesis. a The tube formation activity of HUVECs that were treated with control medium, the culture supernatant of MAD-MB-231 cells (control supernatant), the culture supernatant of FOXP3-overexpressing MAD-MB-231 cells (FOXP3 supernatant), or the culture supernatant of FOXP3-overexpressing MAD-MB-231 cells supplemented with VEGF (FOXP3 supernatant + VEGF). Scale bar, 50 μm. b Quantitation of HUVEC tubulogenesis in the different groups in ( a ). HUVEC spheroid sprouting activity was determined by 3D culture. c Representative images of spheroid sprouting. Scale bar, 100 μm. d Quantitation of the cumulative sprout length (CSL) in different groups. b , d ANOVA with Tukey’s post hoc test

    Techniques Used: Inhibition, Activity Assay, Quantitation Assay

    Inverse correlation between FOXP3 and VEGF expression in human breast cancer samples. a Representative immunohistochemical images of nuclear FOXP3 and VEGF expression in breast cancer specimens. Scale bar, 100 μm (×10) and 20 μm (×40). b A significant negative correlation between nuclear FOXP3 and VEGF expression was found in the breast cancer specimens. b Spearman test
    Figure Legend Snippet: Inverse correlation between FOXP3 and VEGF expression in human breast cancer samples. a Representative immunohistochemical images of nuclear FOXP3 and VEGF expression in breast cancer specimens. Scale bar, 100 μm (×10) and 20 μm (×40). b A significant negative correlation between nuclear FOXP3 and VEGF expression was found in the breast cancer specimens. b Spearman test

    Techniques Used: Expressing, Immunohistochemistry

    FOXP3 downregulates VEGF expression in breast cancer. Real-time PCR was performed to detect the transcription levels of VEGF in breast cancer cell lines with the gain or loss of FOXP3 expression. Transfection of pcDNA3.1-FOXP3 into ( a ) MCF-7, ( b ) T47D or ( c ) MAD-MB-231 cells. Transfection of FOXP3 shRNA into ( d ) MCF-7 or ( e ) T47D cells. Western blotting was performed to detect the expression of VEGF in breast cancer cell lines following the gain or loss of FOXP3 expression. Transfection of pcDNA3.1-FOXP3 into ( f ) MCF-7, ( g ) T47D or ( h ) MAD-MB-231 cells. Transfection of FOXP3 shRNA into ( i ) MCF-7 or ( j ) T47D cells. k Evaluation of FOXP3 and VEGF expression in MAD-MB-231 cells by confocal microscopy (control or FOXP3 overexpression). Scale bar, 20 μm. l Orthotopic injection of MDA-MB-231 cells was performed to generate xenografts, and adenoviruses carrying FOXP3 or control cDNA were injected into the tumors when their volume reached ~50 mm 3 ( n = 5). Serum VEGF levels of mice bearing control or FOXP3-overexpressing tumors were detected by ELISA. a – l Student’s t test
    Figure Legend Snippet: FOXP3 downregulates VEGF expression in breast cancer. Real-time PCR was performed to detect the transcription levels of VEGF in breast cancer cell lines with the gain or loss of FOXP3 expression. Transfection of pcDNA3.1-FOXP3 into ( a ) MCF-7, ( b ) T47D or ( c ) MAD-MB-231 cells. Transfection of FOXP3 shRNA into ( d ) MCF-7 or ( e ) T47D cells. Western blotting was performed to detect the expression of VEGF in breast cancer cell lines following the gain or loss of FOXP3 expression. Transfection of pcDNA3.1-FOXP3 into ( f ) MCF-7, ( g ) T47D or ( h ) MAD-MB-231 cells. Transfection of FOXP3 shRNA into ( i ) MCF-7 or ( j ) T47D cells. k Evaluation of FOXP3 and VEGF expression in MAD-MB-231 cells by confocal microscopy (control or FOXP3 overexpression). Scale bar, 20 μm. l Orthotopic injection of MDA-MB-231 cells was performed to generate xenografts, and adenoviruses carrying FOXP3 or control cDNA were injected into the tumors when their volume reached ~50 mm 3 ( n = 5). Serum VEGF levels of mice bearing control or FOXP3-overexpressing tumors were detected by ELISA. a – l Student’s t test

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Transfection, shRNA, Western Blot, Confocal Microscopy, Over Expression, Injection, Multiple Displacement Amplification, Mouse Assay, Enzyme-linked Immunosorbent Assay

    FOXP3 is a transcriptional suppressor of VEGF. a FOXP3 suppresses VEGF promoter activity, as evaluated by dual-luciferase reporter assays. b The top panel depicts schematic diagrams of the regions amplified by the ChIP primers. The bottom panel shows the amount of DNA precipitated by either the anti-FOXP3 antibody or control IgG; the results are expressed as a percentage of the input genomic DNA from MDA-MB-231 cells. c FOXP3-mediated suppression of the VEGF promoter requires forkhead-binding motifs, as evaluated by dual-luciferase reporter assays. Relative truncation of the VEGF promoter is illustrated in the left panel. Luciferase activity was detected in cells transfected with the truncated VEGF promoter in the right panel. a , c ANOVA with Tukey’s post hoc test
    Figure Legend Snippet: FOXP3 is a transcriptional suppressor of VEGF. a FOXP3 suppresses VEGF promoter activity, as evaluated by dual-luciferase reporter assays. b The top panel depicts schematic diagrams of the regions amplified by the ChIP primers. The bottom panel shows the amount of DNA precipitated by either the anti-FOXP3 antibody or control IgG; the results are expressed as a percentage of the input genomic DNA from MDA-MB-231 cells. c FOXP3-mediated suppression of the VEGF promoter requires forkhead-binding motifs, as evaluated by dual-luciferase reporter assays. Relative truncation of the VEGF promoter is illustrated in the left panel. Luciferase activity was detected in cells transfected with the truncated VEGF promoter in the right panel. a , c ANOVA with Tukey’s post hoc test

    Techniques Used: Activity Assay, Luciferase, Amplification, Chromatin Immunoprecipitation, Multiple Displacement Amplification, Binding Assay, Transfection

    18) Product Images from "Early expressions of hypoxia-inducible factor 1alpha and vascular endothelial growth factor increase the neuronal plasticity of activated endogenous neural stem cells after focal cerebral ischemia"

    Article Title: Early expressions of hypoxia-inducible factor 1alpha and vascular endothelial growth factor increase the neuronal plasticity of activated endogenous neural stem cells after focal cerebral ischemia

    Journal: Neural Regeneration Research

    doi: 10.4103/1673-5374.133136

    Western blot analysis for hypoxia-inducible factor 1α (HIF-1α), vascular endothelial growth factor (VEGF) and nestin expression after focal cerebral ischemia. (A) Representative time course of nestin, HIF-1α, VEGF, and nestin expression in control (sham) and ipsilateral cerebral hemisphere extracts from rats at baseline, 1 h, 12 h, 1 d, 3 d, and 7 d. (B) Time courses of normalized optical density of nestin immunoblots in ipsilateral hemispheres (* P
    Figure Legend Snippet: Western blot analysis for hypoxia-inducible factor 1α (HIF-1α), vascular endothelial growth factor (VEGF) and nestin expression after focal cerebral ischemia. (A) Representative time course of nestin, HIF-1α, VEGF, and nestin expression in control (sham) and ipsilateral cerebral hemisphere extracts from rats at baseline, 1 h, 12 h, 1 d, 3 d, and 7 d. (B) Time courses of normalized optical density of nestin immunoblots in ipsilateral hemispheres (* P

    Techniques Used: Western Blot, Expressing

    19) Product Images from "Promotive effects of bone morphogenetic protein 2 on angiogenesis in hepatocarcinoma via multiple signal pathways"

    Article Title: Promotive effects of bone morphogenetic protein 2 on angiogenesis in hepatocarcinoma via multiple signal pathways

    Journal: Scientific Reports

    doi: 10.1038/srep37499

    Expressions of VEGF, p-P38, p-ERK, p-AKT and p-m-TOR detected by Western blot. ( A ) BMP2 silence down-regulates the expression of VEGF, p-P38, p-ERK, p-AKT and p-m-TOR expressions ( B ) The protein expressions of VEGF, p-P38, p-ERK, p-AKT and p-m-TOR are up-regulated in overexpression-LV-BMP2 group Notes: BMP-2: Bone morphogenetic protein 2; VEGF: vascular endothelial growth factor * Indicates P
    Figure Legend Snippet: Expressions of VEGF, p-P38, p-ERK, p-AKT and p-m-TOR detected by Western blot. ( A ) BMP2 silence down-regulates the expression of VEGF, p-P38, p-ERK, p-AKT and p-m-TOR expressions ( B ) The protein expressions of VEGF, p-P38, p-ERK, p-AKT and p-m-TOR are up-regulated in overexpression-LV-BMP2 group Notes: BMP-2: Bone morphogenetic protein 2; VEGF: vascular endothelial growth factor * Indicates P

    Techniques Used: Western Blot, Expressing, Over Expression

    Related Articles

    Incubation:

    Article Title: Ethanol Promotes New Vessel Growth in Remote Non-Ischemic Myocardium
    Article Snippet: Forty micrograms of whole-tissue lysates were fractionated by SDS-PAGE 4-12% Bis-Tris gels (NuPage Novex Mini Gel, Invitrogen, Carlsbad, CA) and transferred to polyvinylidene difluoride membranes (Millipore, Billerica, MA). .. Membranes were incubated overnight at 4°C with primary antibodies at dilutions recommended by the manufacturer against phosphorylated-endothelial nitric oxide synthase (p-ENOS S1177), ENOS, vascular endothelial growth factor (VEGF), VEGF-receptor 1 (VEGFR1), VEGFR2, vascular endothelial cadherin (VECADHERIN), heat shock protein 27 (HSP27), HSP90, phosphorylated p38 (PP38 T180/Y182), p38, phosphorylated nuclear factor kappa-light-chain-enhancer of activated B cells (pNFκB S536), NFκB, (all from Cell Signaling, Danvers, MA), NADPH oxidase2 (NOX2 Sigma Aldrich, St Louis, MO), endostatin (Millipore), angiostatin (Calbiochem, Billerica, MA), and VEGFR3 (Abcam, Cambridge, MA). .. Membranes were incubated with the appropriate horseradish peroxidase-linked secondary antibody for one hour at room temperature (Jackson ImmunoResearch, West Grove, PA).

    Western Blot:

    Article Title: Obesity but not high-fat diet impairs lymphatic function
    Article Snippet: Cell signal intensity was quantified using MetaMorph (Molecular Devices, Sunnyvale, CA, USA). .. Western blot analysis for VEGFR-3 (Abcam), p-AKT and p-eNOS (Cell Signaling Technology) was performed with total cellular protein harvested from LECs and quantified with NIH Image J. .. Cellular apoptosis and viability assays Apoptosis in response to SA (0.1, 1, 10, 100 μm ) with/without 0.3 nm Phosphatase and tensin homolog inhibitor (PTENi; SF1670, Sigma-Aldrich) was quantified using caspase-3 assay (R & D Systems, Minneapolis, MN, USA) and Annexin V-FITC Apoptosis Detection Kit (eBioscience, San Diego, CA, USA).

    Article Title: The antitumor effect of the novel cancer-specific adenovirus Ad-VEGFR on bladder cancer in vitro and in vivo
    Article Snippet: The recombinant adenovirus vAd-VEGFR-3, which has infective stability, was packed in 293 cells. .. The protein expression level of VEGFR-3 was validated by western blot with an anti-VEGFR antibody (Abcam, Cambridge, Massachusetts, USA). .. EJ cell lines were infected with recombinant adenoviruses vAd-VEGFR-3 or an empty adenovirus vector at a multiplicity of infection (MOI) of 10 pfu/cell for 12 h. The expression of the target gene was tested by qRT-PCR.

    Immunoprecipitation:

    Article Title: Novel Vascular Endothelial Growth Factor D Variants with Increased Biological Activity *
    Article Snippet: After the detection of phosphorylated forms, the membranes were stripped for 15 min at room temperature with Restore Western blot Stripping Buffer (Pierce), and the total amount of protein was detected. .. Total VEGFR-3 was immunoprecipitated with a VEGFR-3 antibody (Abcam). .. The samples were separated using SDS-PAGE and immunoblotted with anti-phosphotyrosine (Upstate) and VEGFR-3 antibodies and detected using a chemiluminescent-based detection system as above.

    Blocking Assay:

    Article Title: LRG1 promotes corneal angiogenesis and lymphangiogenesis in a corneal alkali burn mouse model
    Article Snippet: .. Blocking VEGFR-3 suppresses angiogenic sprouting and vascular network formation. ..

    Expressing:

    Article Title: The antitumor effect of the novel cancer-specific adenovirus Ad-VEGFR on bladder cancer in vitro and in vivo
    Article Snippet: The recombinant adenovirus vAd-VEGFR-3, which has infective stability, was packed in 293 cells. .. The protein expression level of VEGFR-3 was validated by western blot with an anti-VEGFR antibody (Abcam, Cambridge, Massachusetts, USA). .. EJ cell lines were infected with recombinant adenoviruses vAd-VEGFR-3 or an empty adenovirus vector at a multiplicity of infection (MOI) of 10 pfu/cell for 12 h. The expression of the target gene was tested by qRT-PCR.

    Recombinant:

    Article Title: Soluble vascular endothelial growth factor receptor 3 is essential for corneal alymphaticity
    Article Snippet: To estimate VEGF-C’s relative affinity for VEGFR-3 and VEGFR-2, a competitive sandwich ELISA was performed. .. Recombinant human VEGF-C (10 µg/mL, H00007424-P01; Abnova) was coated onto 96-well plates overnight, and equal molar amounts of soluble recombinant VEGFR-3 (CRF113B, Cell Sciences) and recombinant human VEGFR2/KDR/Flk1Fc chimera (R & D; 357KD) were added to each well for the competitive binding to VEGF-C. Antibodies to VEGFR-2 (ab51873, Abcam) and VEGFR-3 (SC-20734, Santa Cruz) were added for 2 hours. .. The wells were washed and HRP-labeled antibodies (antibody to rat IgG, H & L [HRP], ab6734, Abcam, 1:5,000; anti–rabbit IgG, H & L [HRP], NA 934, GE Healthcare) were added for 2 hours.

    Article Title: Hyperoxia sensitizes hypoxic HeLa cells to ionizing radiation by downregulating HIF-1α and VEGF expression
    Article Snippet: Western blotting or IR (feVEGF + IR) were conducted at 48 h post-transfection. .. VEGF/VEGFR stimulation and neutralization tests For the VEGF stimulation test, 10 ng/ml recombinant human VEGF165 (reVEGF, cat. no. ab9571; Abcam) ( ) was added to cell culture media (37°C, 5% CO2 ) at 2 h prior to IR (reVEGF + IR). .. For the ligand-receptor neutralization test, 25 µg/ml VEGF, 30 µg/ml VEGFR1 or 500 ng/ml VEGFR2 neutralization antibodies (cat. nos.

    Binding Assay:

    Article Title: Soluble vascular endothelial growth factor receptor 3 is essential for corneal alymphaticity
    Article Snippet: To estimate VEGF-C’s relative affinity for VEGFR-3 and VEGFR-2, a competitive sandwich ELISA was performed. .. Recombinant human VEGF-C (10 µg/mL, H00007424-P01; Abnova) was coated onto 96-well plates overnight, and equal molar amounts of soluble recombinant VEGFR-3 (CRF113B, Cell Sciences) and recombinant human VEGFR2/KDR/Flk1Fc chimera (R & D; 357KD) were added to each well for the competitive binding to VEGF-C. Antibodies to VEGFR-2 (ab51873, Abcam) and VEGFR-3 (SC-20734, Santa Cruz) were added for 2 hours. .. The wells were washed and HRP-labeled antibodies (antibody to rat IgG, H & L [HRP], ab6734, Abcam, 1:5,000; anti–rabbit IgG, H & L [HRP], NA 934, GE Healthcare) were added for 2 hours.

    Neutralization:

    Article Title: Hyperoxia sensitizes hypoxic HeLa cells to ionizing radiation by downregulating HIF-1α and VEGF expression
    Article Snippet: Western blotting or IR (feVEGF + IR) were conducted at 48 h post-transfection. .. VEGF/VEGFR stimulation and neutralization tests For the VEGF stimulation test, 10 ng/ml recombinant human VEGF165 (reVEGF, cat. no. ab9571; Abcam) ( ) was added to cell culture media (37°C, 5% CO2 ) at 2 h prior to IR (reVEGF + IR). .. For the ligand-receptor neutralization test, 25 µg/ml VEGF, 30 µg/ml VEGFR1 or 500 ng/ml VEGFR2 neutralization antibodies (cat. nos.

    Cell Culture:

    Article Title: Hyperoxia sensitizes hypoxic HeLa cells to ionizing radiation by downregulating HIF-1α and VEGF expression
    Article Snippet: Western blotting or IR (feVEGF + IR) were conducted at 48 h post-transfection. .. VEGF/VEGFR stimulation and neutralization tests For the VEGF stimulation test, 10 ng/ml recombinant human VEGF165 (reVEGF, cat. no. ab9571; Abcam) ( ) was added to cell culture media (37°C, 5% CO2 ) at 2 h prior to IR (reVEGF + IR). .. For the ligand-receptor neutralization test, 25 µg/ml VEGF, 30 µg/ml VEGFR1 or 500 ng/ml VEGFR2 neutralization antibodies (cat. nos.

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    Abcam anti vegf a
    Conformer selectivity of the peptides for different oncogenic quadruplexes in MCF-7 cells. ( A ) pGL4.72[ hRlucCP ] vector having the inserts containing oncogenic promoter sequences ( c-MYC, BCL-2, KRAS and <t>VEGF-A</t> ) and upstream G-quadruplex forming elements ahead of the hRluc coding region. The promoter sequences are cloned into KpnI and HindIII restriction sites with or without the wild-type quadruplex scaffolds. hRluc, Renilla luciferase gene; hCL1 and hPEST , protein destabilizing sequences; oriC, origin of replication; AmpR, ampicillin resistance gene; SV40 (Simian virus 40 polyadenylation signal cassette), P1 and P2, promoter sequences; null, constructs having no quadruplex motif upstream the oncogene promoters; GQ, G-quadruplex forming motif. ( B ) Dual-luciferase assays. Evaluation of the promoter activity of different oncogenes (( B ) c-MYC , ( C ) BCL-2 , ( D ) KRAS and ( E ) VEGF-A ) of using the reporter plasmids with or without the wild-type quadruplex-forming sequences (Pu27-GQ, BCL-2-GQ, KRAS-GQ and VEGF-A-GQ) in MCF-7 cells. Relative promoter activity is determined by normalizing the Rluc/Fluc values to that of the cells transfected with P1-P2 promoter construct (GQ-null), having no quadruplex-forming motif. Error bars represent mean ± SE ( N = 5). Statistical differences in the luciferase activities compared to the respective GQ-null constructs of the target oncogenes used one-way ANOVA followed by Tukey–Kramer Test ( # P
    Anti Vegf A, supplied by Abcam, 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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    vegf  (Abcam)
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    Abcam vegf
    RT-qPCR and Western blot analysis demonstrated that MBNL1-AS1 reduced the expression of KCNMA1 and cGMP-PKG signaling pathway-related genes. a RT-qPCR measured the relative mRNA expression of MBNL1-AS1, KCNMA1, PKGII, <t>VASP,</t> <t>VEGF,</t> Bax, and Bcl-2 in skeletal muscle cells after transfection. b Protein bands measured by Western blot analysis. c Western blot analysis measured the relative protein levels of KCNMA1, PKGII, VASP, p-PKGII, p-VASP, VEGF, Bax, and Bcl-2 in skeletal muscle cells after transfection. MBNL1-AS1 muscleblind-like 1 antisense RNA 1, KCNMA1 potassium calcium-activated channel subfamily M alpha 1, PKGII type II cyclic guanosine monophosphate-dependent protein kinase G, p-PKGII phosphorylated-PKGII, VASP vasodilator-stimulated phosphoprotein, p-VASP phosphorylated-VASP, VEGF vascular endothelial growth factor, Bcl-2 B-cell lymphoma/leukemia-2, Bax Bcl-2 associated X protein, si small interfering, GAPDH glyceraldehyde-3-phosphate dehydrogenase, NC negative control; * p
    Vegf, supplied by Abcam, 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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    Abcam rabbit anti vegf a
    Lack of p73 affects TGF- β signaling in vivo . Quantification of <t>VEGF-A</t> expression and analysis of the TGF -β /ALK1/ID1 signaling pathway in P5 retinas from WT and p73KO mice. qRT-PCR analysis demonstrated a significant decrease in VEGF-A
    Rabbit Anti Vegf A, supplied by Abcam, 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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    Conformer selectivity of the peptides for different oncogenic quadruplexes in MCF-7 cells. ( A ) pGL4.72[ hRlucCP ] vector having the inserts containing oncogenic promoter sequences ( c-MYC, BCL-2, KRAS and VEGF-A ) and upstream G-quadruplex forming elements ahead of the hRluc coding region. The promoter sequences are cloned into KpnI and HindIII restriction sites with or without the wild-type quadruplex scaffolds. hRluc, Renilla luciferase gene; hCL1 and hPEST , protein destabilizing sequences; oriC, origin of replication; AmpR, ampicillin resistance gene; SV40 (Simian virus 40 polyadenylation signal cassette), P1 and P2, promoter sequences; null, constructs having no quadruplex motif upstream the oncogene promoters; GQ, G-quadruplex forming motif. ( B ) Dual-luciferase assays. Evaluation of the promoter activity of different oncogenes (( B ) c-MYC , ( C ) BCL-2 , ( D ) KRAS and ( E ) VEGF-A ) of using the reporter plasmids with or without the wild-type quadruplex-forming sequences (Pu27-GQ, BCL-2-GQ, KRAS-GQ and VEGF-A-GQ) in MCF-7 cells. Relative promoter activity is determined by normalizing the Rluc/Fluc values to that of the cells transfected with P1-P2 promoter construct (GQ-null), having no quadruplex-forming motif. Error bars represent mean ± SE ( N = 5). Statistical differences in the luciferase activities compared to the respective GQ-null constructs of the target oncogenes used one-way ANOVA followed by Tukey–Kramer Test ( # P

    Journal: Nucleic Acids Research

    Article Title: Site-specific amino acid substitution in dodecameric peptides determines the stability and unfolding of c-MYC quadruplex promoting apoptosis in cancer cells

    doi: 10.1093/nar/gky824

    Figure Lengend Snippet: Conformer selectivity of the peptides for different oncogenic quadruplexes in MCF-7 cells. ( A ) pGL4.72[ hRlucCP ] vector having the inserts containing oncogenic promoter sequences ( c-MYC, BCL-2, KRAS and VEGF-A ) and upstream G-quadruplex forming elements ahead of the hRluc coding region. The promoter sequences are cloned into KpnI and HindIII restriction sites with or without the wild-type quadruplex scaffolds. hRluc, Renilla luciferase gene; hCL1 and hPEST , protein destabilizing sequences; oriC, origin of replication; AmpR, ampicillin resistance gene; SV40 (Simian virus 40 polyadenylation signal cassette), P1 and P2, promoter sequences; null, constructs having no quadruplex motif upstream the oncogene promoters; GQ, G-quadruplex forming motif. ( B ) Dual-luciferase assays. Evaluation of the promoter activity of different oncogenes (( B ) c-MYC , ( C ) BCL-2 , ( D ) KRAS and ( E ) VEGF-A ) of using the reporter plasmids with or without the wild-type quadruplex-forming sequences (Pu27-GQ, BCL-2-GQ, KRAS-GQ and VEGF-A-GQ) in MCF-7 cells. Relative promoter activity is determined by normalizing the Rluc/Fluc values to that of the cells transfected with P1-P2 promoter construct (GQ-null), having no quadruplex-forming motif. Error bars represent mean ± SE ( N = 5). Statistical differences in the luciferase activities compared to the respective GQ-null constructs of the target oncogenes used one-way ANOVA followed by Tukey–Kramer Test ( # P

    Article Snippet: Santacruz Biotechnology Inc.), anti E2F1, anti VEGF-A, anti-P53, anti-APAF1, anti-Caspase 8, anti-PARP (Abcam), anti-β-actin (Mouse monoclonal, 1:10 000 dil.

    Techniques: Plasmid Preparation, Clone Assay, Luciferase, Construct, Activity Assay, Transfection

    Selective arresting of c-MYC quadruplex by KR12C induce apoptotic signalling in cancer cells. ( A ) Western blot analyses of the major apoptotic markers (BCL-2, APAF-1, caspase 8 and PARP). ( B ) Estimation of protein expression level of the apoptotic markers by semi-densitometric analyses. Estimation of BCL-2 transcripts by real time PCR analyses at 5 and 10 μM KR12C concentration. ( C ) Western blot analyses of other proteins associated with the signalling cascade (c-MYC, E2F-1, VEGF-A and P53). ( D ) Quatification of protein and mRNA expression level of c-MYC, E2F-1, VEGF-A and P53 by semi-densitometric analyses and real time PCR respectively. Arrow pointing upwards and downwards denote the upregulation of the respective protein and mRNA expression level. Error bars represent mean ± SE ( N = 3). Statistical differences are determined compared to the control by two-tailed Student's t test (* P

    Journal: Nucleic Acids Research

    Article Title: Site-specific amino acid substitution in dodecameric peptides determines the stability and unfolding of c-MYC quadruplex promoting apoptosis in cancer cells

    doi: 10.1093/nar/gky824

    Figure Lengend Snippet: Selective arresting of c-MYC quadruplex by KR12C induce apoptotic signalling in cancer cells. ( A ) Western blot analyses of the major apoptotic markers (BCL-2, APAF-1, caspase 8 and PARP). ( B ) Estimation of protein expression level of the apoptotic markers by semi-densitometric analyses. Estimation of BCL-2 transcripts by real time PCR analyses at 5 and 10 μM KR12C concentration. ( C ) Western blot analyses of other proteins associated with the signalling cascade (c-MYC, E2F-1, VEGF-A and P53). ( D ) Quatification of protein and mRNA expression level of c-MYC, E2F-1, VEGF-A and P53 by semi-densitometric analyses and real time PCR respectively. Arrow pointing upwards and downwards denote the upregulation of the respective protein and mRNA expression level. Error bars represent mean ± SE ( N = 3). Statistical differences are determined compared to the control by two-tailed Student's t test (* P

    Article Snippet: Santacruz Biotechnology Inc.), anti E2F1, anti VEGF-A, anti-P53, anti-APAF1, anti-Caspase 8, anti-PARP (Abcam), anti-β-actin (Mouse monoclonal, 1:10 000 dil.

    Techniques: Western Blot, Expressing, Real-time Polymerase Chain Reaction, Concentration Assay, Two Tailed Test

    Schematic representation showing selective quadruplex interaction at c-MYC promoter by KR12C promotes apoptotic signalling in VEGF-A-BCL-2 axis in MCF-7 cells.

    Journal: Nucleic Acids Research

    Article Title: Site-specific amino acid substitution in dodecameric peptides determines the stability and unfolding of c-MYC quadruplex promoting apoptosis in cancer cells

    doi: 10.1093/nar/gky824

    Figure Lengend Snippet: Schematic representation showing selective quadruplex interaction at c-MYC promoter by KR12C promotes apoptotic signalling in VEGF-A-BCL-2 axis in MCF-7 cells.

    Article Snippet: Santacruz Biotechnology Inc.), anti E2F1, anti VEGF-A, anti-P53, anti-APAF1, anti-Caspase 8, anti-PARP (Abcam), anti-β-actin (Mouse monoclonal, 1:10 000 dil.

    Techniques:

    RT-qPCR and Western blot analysis demonstrated that MBNL1-AS1 reduced the expression of KCNMA1 and cGMP-PKG signaling pathway-related genes. a RT-qPCR measured the relative mRNA expression of MBNL1-AS1, KCNMA1, PKGII, VASP, VEGF, Bax, and Bcl-2 in skeletal muscle cells after transfection. b Protein bands measured by Western blot analysis. c Western blot analysis measured the relative protein levels of KCNMA1, PKGII, VASP, p-PKGII, p-VASP, VEGF, Bax, and Bcl-2 in skeletal muscle cells after transfection. MBNL1-AS1 muscleblind-like 1 antisense RNA 1, KCNMA1 potassium calcium-activated channel subfamily M alpha 1, PKGII type II cyclic guanosine monophosphate-dependent protein kinase G, p-PKGII phosphorylated-PKGII, VASP vasodilator-stimulated phosphoprotein, p-VASP phosphorylated-VASP, VEGF vascular endothelial growth factor, Bcl-2 B-cell lymphoma/leukemia-2, Bax Bcl-2 associated X protein, si small interfering, GAPDH glyceraldehyde-3-phosphate dehydrogenase, NC negative control; * p

    Journal: Experimental & Molecular Medicine

    Article Title: Downregulation of the long noncoding RNA MBNL1-AS1 protects sevoflurane-pretreated mice against ischemia-reperfusion injury by targeting KCNMA1

    doi: 10.1038/s12276-018-0133-y

    Figure Lengend Snippet: RT-qPCR and Western blot analysis demonstrated that MBNL1-AS1 reduced the expression of KCNMA1 and cGMP-PKG signaling pathway-related genes. a RT-qPCR measured the relative mRNA expression of MBNL1-AS1, KCNMA1, PKGII, VASP, VEGF, Bax, and Bcl-2 in skeletal muscle cells after transfection. b Protein bands measured by Western blot analysis. c Western blot analysis measured the relative protein levels of KCNMA1, PKGII, VASP, p-PKGII, p-VASP, VEGF, Bax, and Bcl-2 in skeletal muscle cells after transfection. MBNL1-AS1 muscleblind-like 1 antisense RNA 1, KCNMA1 potassium calcium-activated channel subfamily M alpha 1, PKGII type II cyclic guanosine monophosphate-dependent protein kinase G, p-PKGII phosphorylated-PKGII, VASP vasodilator-stimulated phosphoprotein, p-VASP phosphorylated-VASP, VEGF vascular endothelial growth factor, Bcl-2 B-cell lymphoma/leukemia-2, Bax Bcl-2 associated X protein, si small interfering, GAPDH glyceraldehyde-3-phosphate dehydrogenase, NC negative control; * p

    Article Snippet: The membrane was then blocked in 5% skimmed milk at room temperature for 1 h, washed twice with PBS, and then incubated overnight at 4 °C with diluted rabbit anti-mouse primary antibodies against KCNMA1 (1:1000, MAB8589; AmyJet Scientific Inc., Wuhan, China), PKGII (1:1000, ab145063; Abcam, Inc., Cambridge, MA, USA), VASP (1:1000, ab58555; Abcam, USA), VEGF (1:1000, ab32152; Abcam, USA), Bax (1:1000, ab32503; Abcam, USA), Bcl-2 (1:1000, ab59348; Abcam, USA), Cyclin D1 (1:1000, ab134175, Abcam, USA), Cyclin D3 (1:1000, ab28283, Abcam, USA), Cdc 42 (1:1000, ab187643, Abcam, USA), caspase-3 (1:1000, ab208161, Abcam, USA), and PARP (1:1000, ab32064, Abcam, USA).

    Techniques: Quantitative RT-PCR, Western Blot, Expressing, Transfection, Negative Control

    MBNL1-AS1 was highly expressed, but KCNMA1 and cGMP-PKG pathway-related factors were poorly expressed in mice with I/R injury after TKA. a Relative mRNA expression of MBNL1-AS1, KCNMA1, PKGII, VASP, VEGF, Bax, and Bcl-2 in the skeletal muscle tissues of mice detected by RT-qPCR. b Protein bands observed after Western blot analysis. c Relative protein levels of KCNMA1, PKGII, VASP, p-PKGII, p-VASP, VEGF, Bax, and Bcl-2 in skeletal muscle tissues of mice evaluated by Western blot analysis. I/R ischemia-reperfusion, Sevo sevoflurane, MBNL1-AS1 muscleblind-like 1 antisense RNA 1, KCNMA1 potassium calcium-activated channel subfamily alpha 1, PKGII type II cyclic guanosine monophosphate-dependent protein kinase G, VASP vasodilator-stimulated phosphoprotein, VEGF vascular endothelial growth factor, Bcl-2 B-cell lymphoma/leukemia-2, Bax Bcl-2 associated X protein, GAPDH glyceraldehyde-3-phosphate dehydrogenase; * p

    Journal: Experimental & Molecular Medicine

    Article Title: Downregulation of the long noncoding RNA MBNL1-AS1 protects sevoflurane-pretreated mice against ischemia-reperfusion injury by targeting KCNMA1

    doi: 10.1038/s12276-018-0133-y

    Figure Lengend Snippet: MBNL1-AS1 was highly expressed, but KCNMA1 and cGMP-PKG pathway-related factors were poorly expressed in mice with I/R injury after TKA. a Relative mRNA expression of MBNL1-AS1, KCNMA1, PKGII, VASP, VEGF, Bax, and Bcl-2 in the skeletal muscle tissues of mice detected by RT-qPCR. b Protein bands observed after Western blot analysis. c Relative protein levels of KCNMA1, PKGII, VASP, p-PKGII, p-VASP, VEGF, Bax, and Bcl-2 in skeletal muscle tissues of mice evaluated by Western blot analysis. I/R ischemia-reperfusion, Sevo sevoflurane, MBNL1-AS1 muscleblind-like 1 antisense RNA 1, KCNMA1 potassium calcium-activated channel subfamily alpha 1, PKGII type II cyclic guanosine monophosphate-dependent protein kinase G, VASP vasodilator-stimulated phosphoprotein, VEGF vascular endothelial growth factor, Bcl-2 B-cell lymphoma/leukemia-2, Bax Bcl-2 associated X protein, GAPDH glyceraldehyde-3-phosphate dehydrogenase; * p

    Article Snippet: The membrane was then blocked in 5% skimmed milk at room temperature for 1 h, washed twice with PBS, and then incubated overnight at 4 °C with diluted rabbit anti-mouse primary antibodies against KCNMA1 (1:1000, MAB8589; AmyJet Scientific Inc., Wuhan, China), PKGII (1:1000, ab145063; Abcam, Inc., Cambridge, MA, USA), VASP (1:1000, ab58555; Abcam, USA), VEGF (1:1000, ab32152; Abcam, USA), Bax (1:1000, ab32503; Abcam, USA), Bcl-2 (1:1000, ab59348; Abcam, USA), Cyclin D1 (1:1000, ab134175, Abcam, USA), Cyclin D3 (1:1000, ab28283, Abcam, USA), Cdc 42 (1:1000, ab187643, Abcam, USA), caspase-3 (1:1000, ab208161, Abcam, USA), and PARP (1:1000, ab32064, Abcam, USA).

    Techniques: Mouse Assay, Expressing, Quantitative RT-PCR, Western Blot

    VEGF-A and VEGFR-1 are important for Snail expression. ( a ) Cells were transfected with indicated vectors. Quantitative RT-PCR (mean±s.d. * P

    Journal: Experimental & Molecular Medicine

    Article Title: Rab25 augments cancer cell invasiveness through a β1 integrin/EGFR/VEGF-A/Snail signaling axis and expression of fascin

    doi: 10.1038/emm.2017.248

    Figure Lengend Snippet: VEGF-A and VEGFR-1 are important for Snail expression. ( a ) Cells were transfected with indicated vectors. Quantitative RT-PCR (mean±s.d. * P

    Article Snippet: Immunohistochemistry was performed with primary Rab25 (ab106175, 1:200, Abcam, Cambridge, MA, USA), VEGFR-1 (ab32152, 1:250, Abcam), Snail (ab180714, 1:100, Abcam) and fascin (ab126772, 1:250, Abcam) antibodies, as described previously.

    Techniques: Expressing, Transfection, Quantitative RT-PCR

    Lack of p73 affects TGF- β signaling in vivo . Quantification of VEGF-A expression and analysis of the TGF -β /ALK1/ID1 signaling pathway in P5 retinas from WT and p73KO mice. qRT-PCR analysis demonstrated a significant decrease in VEGF-A

    Journal: Cell Death and Differentiation

    Article Title: p73 is required for endothelial cell differentiation, migration and the formation of vascular networks regulating VEGF and TGFβ signaling

    doi: 10.1038/cdd.2014.214

    Figure Lengend Snippet: Lack of p73 affects TGF- β signaling in vivo . Quantification of VEGF-A expression and analysis of the TGF -β /ALK1/ID1 signaling pathway in P5 retinas from WT and p73KO mice. qRT-PCR analysis demonstrated a significant decrease in VEGF-A

    Article Snippet: Western blotting was performed as previously described with the following primary antibodies: rabbit anti-pSmad1/5 (Ser463/465) 1 : 1000 (Cell Signaling, Danvers, MA, USA), rabbit anti Smad1 1 : 1000 (Cell Signaling), mouse anti-pERK 1 : 1000 (Santa Cruz Biotechnology), rabbit anti-ERK1 : 10 000 (Santa Cruz Biotechnology), rabbit anti-VEGF-A 1 : 1000 (Abcam) mouse anti DNp73 1 : 1000 (Imgenex, San Diego, CO, USA), and rabbit anti Nanog 1 : 1000 (Chemicon, Billerica, TX, USA) followed by the appropriate HRP-conjugated secondary antibodies (Pierce, Waltham, MA, USA).

    Techniques: In Vivo, Expressing, Mouse Assay, Quantitative RT-PCR