vascular endothelial growth factor vegf  (Santa Cruz Biotechnology)

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Production of Vascular Endothelial Growth Factors from Human Lung Macrophages Induced by Group IIA and Group X Secreted Phospholipases A2

doi: 10.4049/jimmunol.0902501

Figure Lengend Snippet: sPLA 2 s induce the release of VEGF-A and VEGF-C from HLMs. A and B , HLMs were incubated (37°C, 24 h) with RPMI 1640 alone (Control) or with the indicated concentrations of hGIIA, hGX, or LPS. VEGF-A ( A ) and VEGF-C ( B ) release was determined by ELISA. Data are mean ± SEM of four experiments. * p

Article Snippet: The following were purchased: LPS (from Escherichia coli serotype 026:B6), 5′-(N-ethylcarboxamido)-adenosine (NECA), 2- p -(2-Carboxyethyl)phenethylamino-5′-N-ethylcarboxamidoadenosine , 2-Chloro-N6 -(3-iodobenzyl)-adenosine-5′-N-methyluronamide (Cl-IB-MECA), Pipes, BSA, Percoll, l -glutamine, antibiotic-antimycotic solution (10,000 UI/ml penicillin, 10 mg/ml streptomycin, and 25 μg/ml amphotericin B), Triton X-100, Polymyxin B (Sigma-Aldrich, St. Louis, MO); RPMI 1640, FCS, and guanidine hydrochloride (MP Biomedicals Europe, Illkirch, France); rabbit polyclonal anti–VEGF-B (H-70), anti–VEGF-C (H-190), and anti–VEGF-D (H-144) Abs, goat polyclonal anti-PlGF (C-20) and anti-GAPDH (V-18) Abs, normal rabbit and goat IgG Abs, HRP-conjugated donkey anti-rabbit and anti-goat IgG Abs (Santa Cruz Biotechnology, Santa Cruz, CA); goat polyclonal anti–VEGF-A Ab, human recombinant VEGF-A165 , and VEGF-A121 (R & D System, Minneapolis, MN); SB203580 (Calbiochem, La Jolla, CA); PD98059 (Cell Signaling, Beverly, MA).

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Production of Vascular Endothelial Growth Factors from Human Lung Macrophages Induced by Group IIA and Group X Secreted Phospholipases A2

doi: 10.4049/jimmunol.0902501

Figure Lengend Snippet: HLMs constitutively express different forms of VEGF. A , Expression of VEGF mRNAs. RNA extraction from resting HLMs and RT-PCR was performed as described under Materials and Methods . Specific RT-PCR amplification products for VEGFA (isoforms 189, 165, and 121), VEGFB (isoforms 186 and 167), VEGFC, VEGFD, PlGF , and GAPDH were separated on 2% agarose gel, stained with ethidium bromide, and visualized with an image analysis system. The experiment shown is representative of three separate experiments. B , Detection of VEGF proteins. HLM protein extracts (40 μg per sample) were immunoblotted with anti–VEGF-A (gel I and II), anti-PlGF (gel III), anti–VEGF-B (gel IV), anti–VEGF-C (gel V), and anti–VEGF-D (gel VI) Abs. rhVEGF-A 165 , MCF-7 cells, EBNA expressing PlGF-1, RAW 264.7 cells were used as positive controls. Stripped membranes were reprobed with anti-GAPDH Ab to confirm equal protein content of each sample. Each Western blot shown is representative of three separate experiments.

Article Snippet: The following were purchased: LPS (from Escherichia coli serotype 026:B6), 5′-(N-ethylcarboxamido)-adenosine (NECA), 2- p -(2-Carboxyethyl)phenethylamino-5′-N-ethylcarboxamidoadenosine , 2-Chloro-N6 -(3-iodobenzyl)-adenosine-5′-N-methyluronamide (Cl-IB-MECA), Pipes, BSA, Percoll, l -glutamine, antibiotic-antimycotic solution (10,000 UI/ml penicillin, 10 mg/ml streptomycin, and 25 μg/ml amphotericin B), Triton X-100, Polymyxin B (Sigma-Aldrich, St. Louis, MO); RPMI 1640, FCS, and guanidine hydrochloride (MP Biomedicals Europe, Illkirch, France); rabbit polyclonal anti–VEGF-B (H-70), anti–VEGF-C (H-190), and anti–VEGF-D (H-144) Abs, goat polyclonal anti-PlGF (C-20) and anti-GAPDH (V-18) Abs, normal rabbit and goat IgG Abs, HRP-conjugated donkey anti-rabbit and anti-goat IgG Abs (Santa Cruz Biotechnology, Santa Cruz, CA); goat polyclonal anti–VEGF-A Ab, human recombinant VEGF-A165 , and VEGF-A121 (R & D System, Minneapolis, MN); SB203580 (Calbiochem, La Jolla, CA); PD98059 (Cell Signaling, Beverly, MA).

Techniques: Expressing, RNA Extraction, Reverse Transcription Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis, Staining, Western Blot

Journal: PLoS ONE

Article Title: Cyclin A1 Modulates the Expression of Vascular Endothelial Growth Factor and Promotes Hormone-Dependent Growth and Angiogenesis of Breast Cancer

doi: 10.1371/journal.pone.0072210

Figure Lengend Snippet: Effect of cyclin A1 expression on tumor invasion is associated with its effect on VEGF expression in MCF-7 cells. (A) Evaluation of cyclin A1 and VEGF expression in metastatic lesions from lymph nodes from patients with breast cancer metastasis using immunohistochemical analysis. Representative pictures show the cancer cells are strongly positive to cyclin A1 and VEGF expression. Upper panels represent cores at 20x magnificantion and lower panels represent the higher magnification (40x) of the selected areas. (B) MCF-7 cells that were transfected with cyclin A1pCMS-EGFP or pCMS-EGFP vectors were applied on the Matrigel-coated invasion chamber and were assessed after 48 or 72 hours. Data in graphs are the mean ± SD represents two independent experiments, each performed in duplicates. P value is indicated. (C) MDA-MB-231 cells transfected with cyclin A1pCMS-EGFP or pCMS-EGFP were applied on the Matrigel-coated invasion chamber and were analyzed after 48 or 72 hours. Data in graphs are the mean of two independent experiments, each performed in duplicate, p=0.002 for 48 h and p=0.02 for 72 h. (D) Cell cycle distribution of the cells that were transfected with cyclin A1pCMS-EGFP or pCMS-EGFP. Data in graphs are the mean ± SD represents three independent experiments from flow cytometry analysis. The percentage of cells at onset of each cell cycle phase is indicated. (E) Western blot analysis shows the levels of cyclin D1 and CDK1 protein expression in the cells that were transfected with cyclin A1pCMS-EGFP or pCMS-EGFP. (F) Representative picture shows the VEGF mRNA expression in the cells transfected with cyclin A1pCMS-EGFP or pCMS-EGFP (upper panel). Quantification of VEGF mRNA level in the samples is indicated. P value is shown (lower panel). (G) ELISA assay of VEGF secretion in the cells transfected with cyclin A1pCMS-EGFP or pCMS-EGFP. Mean ± SD represents three independent experiments (lower panel). Breast cancer cell lines used for these studies are T47D, MCF-7 and MDA-MB231 as indicated.

Article Snippet: Polyclonal human anti-CD31 and CDK1 (BD Pharmingen), anti-β-actin, anti-VEGF, anti-VEGF-R1 and VEGF-R2 (Santa Cruz Biotechnology, CA), anti-ER-α, Ki67 (Dako, Golstrup, Denmark) and anti-cyclin D1 (Cell Signaling Technology Inc, Danvers, MA).

Techniques: Expressing, Immunohistochemistry, Transfection, Multiple Displacement Amplification, Flow Cytometry, Cytometry, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Cyclin A1 Modulates the Expression of Vascular Endothelial Growth Factor and Promotes Hormone-Dependent Growth and Angiogenesis of Breast Cancer

doi: 10.1371/journal.pone.0072210

Figure Lengend Snippet: Long-term effect of elevated level of cyclin A1 on growth and angiogenesis phenotype of xenograft tumors in mice. MCF-7 cells stable expressing pcDNA–cyclin A1 or pcDNA vectors were subcutaneous implanted into female nude mice with E2 supplementation. (A, B) Representative microphotographs of xenograft tumor sections stained with Haematoxylin and Eosin are shown. (C, D) Representative pictures show the xenograft tumors stained with antibody against human CD31, the CD31 positive vessels are indicated. The control tumor “control-pcDNA” and cyclin A1 expressing tumors “cyclin A1-pcDNA” are indicated. (E) Growth curves of the two groups of xenograft tumors are indicated. The time is indicated in x-axis and tumor volume in mm 3 is indicated in y-axis. (F) Quantification of the tumor vascularizations in cyclin A1 expressing xenograft tumors “cyclin A1-pcDNA” and in control xenograft tumors “control-pcDNA”. The numbers of CD31-positive blood vessels in the central vs. edge regions of the tumor areas are shown. P values are indicated. Mean ± SD represents three independent experiments. (G–N) Xenograft tumors from “cyclin A1-pcDNA” and “control-pcDNA” groups were immunostained with antibodies against VEGF, VEGFR1, ER-α and Ki67. The representative microphotographs are shown.

Article Snippet: Polyclonal human anti-CD31 and CDK1 (BD Pharmingen), anti-β-actin, anti-VEGF, anti-VEGF-R1 and VEGF-R2 (Santa Cruz Biotechnology, CA), anti-ER-α, Ki67 (Dako, Golstrup, Denmark) and anti-cyclin D1 (Cell Signaling Technology Inc, Danvers, MA).

Techniques: Mouse Assay, Expressing, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Neurons produce FGF2 and VEGF and secrete them at least in part by shedding extracellular vesicles

doi: 10.1111/j.1582-4934.2007.00100.x

Figure Lengend Snippet: Neurons cultured for 15 days in serum-free Maat Medium release extracellular vesicles that contain both VEGF and FGF-2. A ( a ): Typical neuronal culture, observed in bright field, showing many extracellular vesicles of different sizes, some of which are indicated by arrows for reference; A ( b ) immunofluorescent staining of nuclei with DAPI; same field as in A ( a ). B : Schematic drawing of the protocol used to prepare an extracellular vesicle fraction (according to Dolo et al. , 1994). C : Western analysis of total cell lysates (lane 1) and vesicles (lane 2) prepared, as outlined in B, from neurons cultured for 15 days in Maat medium and then labelled with 35 S-methionine for 24 hrs. Proteins were immunostained with either rabbit polyclonal anti-human VEGF antibodies ( a ) or mouse monoclonal anti-FGF-2 ( b ). After electrophoresis and western blot, the same membranes were also exposed to X-ray sensitive films for 3–5 days: an example of the radioactive protein pattern, 48 hrs after metabolic labelling and medium replacement, is shown in ( c ). Arrowheads in ( a ) and arrows in ( b ) indicate VEGF isoforms and FGF-2 isoforms, respectively. The arrow in ( c ) indicates the only band that could be interpreted as an FGF-2 isoform. The arrowhead in ( c ) indicates the only band that should represent a radioactive VEGF isoform.

Article Snippet: Cells were finally incubated with rabbit polyclonal anti-human VEGF165 antibodies (Calbiochem) that recognize all the known VEGF isoforms, mouse monoclonal anti-bovine purified FGF-2 antobodies (Upstate) that recognize all the known isoforms of FGF-2, and goat anti-integrin β-1 (Santa Cruz, CA, USA).

Techniques: Cell Culture, Staining, Western Blot, Electrophoresis, Metabolic Labelling

Journal: Nature materials

Article Title: Parenchymal and Stromal Tissue Regeneration of Tooth Organ by Pivotal Signals Reinstated in Decellularized Matrix

doi: 10.1038/s41563-019-0368-6

Figure Lengend Snippet: Structural and mechanical properties of regenerated dentin and overall schematic of Alx3/Wnt/VEGF cascades. a: Scanning electron microscopy of dental pulp (p), newly-formed dentin (nd) and native dentin (d) in cross-section. Scale bar: 200 μm. b,c: Dentinal tubules and resin casts in the newly-formed dentin (blue dashed box in a ) and native dentin (yellow dashed box in a ) by SEM. Scale bar: 50 μm. d,e: Odontoblast processes casts in the native dentin ( d,e ) and newly-formed dentin ( f,g ) of a native porcine incisor by SEM. d: dentin; dt: dentinal tubules; OdP: odontoblast process. d,f: scale bar: 50 μm. e,g: scale bar: 10 μm. h: Newly-formed dentin volume; i: Dentin mineral density, j: Ratio of newly-formed dentin (N.D.) and native dentin; k: Harness (MPa); I: Young’s modules (GPa). ( h-j: n=5 independent biological tooth samples; k,l: n=4 independent biological tooth samples; Presented as mean±SD; p values calculated by one-way ANOVA with Bonferroni; * p

Article Snippet: Alx3 conditioned medium was incubated with VEGF (sc-7269, 1 μg per 400 μg total protein, Santa Cruz) at 4°C overnight, with normal goat IgG serum as a negative control.

Techniques: Electron Microscopy

Journal: Nature materials

Article Title: Parenchymal and Stromal Tissue Regeneration of Tooth Organ by Pivotal Signals Reinstated in Decellularized Matrix

doi: 10.1038/s41563-019-0368-6

Figure Lengend Snippet: Alx3-restored adult mesenchyme stem/progenitor cells (MSCs) induced angiogenesis and VEGF signaling. a,a’,d,d’: Angiogenesis in regenerated tissues. a,d: scale bar, 200 μm. a’,d’: scale bar, 100 μm. d: native dentin. b,b’,e,e’: Human von Willebrand factor immunohistochemistry. Green arrowheads: endothelial cells of human origin ( e’ ); black arrowheads: endothelial cells of host (mouse) origin ( e’ ); red arrowheads: chimeric blood vessels ( e’ ). b,e: scale bar, 200 μm. b’,e’: scale bar, 50 μm. c,f: Human mitochondria staining; red arrowheads: human cells; black arrowheads: host (mouse) cells. Scale bar, 100 μm. a-h: n=5 independent biological samples, g: Blood vessel (b.v.) quantification (n=5 independent biological samples; presented as mean±SD; p values calculated by one-way ANOVA with Bonferroni; **: p

Article Snippet: Alx3 conditioned medium was incubated with VEGF (sc-7269, 1 μg per 400 μg total protein, Santa Cruz) at 4°C overnight, with normal goat IgG serum as a negative control.

Techniques: Immunohistochemistry, Staining