vegf Search Results


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  • 96
    Millipore vegf a
    VASH1 inhibits angiogenesis and lymphangiogenesis induced by <t>VEGF-A.</t> A: Pellets containing 160 ng of VEGF-A plus or minus 4 ng of VASH1 were inoculated into the mouse cornea. Fourteen days after the inoculation, the corneas were harvested and immunostained
    Vegf A, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 529 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Santa Cruz Biotechnology vegf
    Effects of MEK-ERK inhibition on <t>VEGF-dependent</t> proliferation and differentiation. ( A ) Western blots of VEGF-induced phospho-ERK. E6 retinas were cultured with or without U0126 for 60 minutes before VEGF was added for 10 minutes. Blots were probed with phospho-ERK1/2 (top) and <t>α-tubulin</t> (bottom) antibodies.( B ) Quantification of optical densities of phospho-ERK signals were normalized against α-tubulin signals ( n =4). ( C ) RT-PCR detection of cyclin D1 transcripts in E6 retinas cultured for 24 hours in the presence or absence of VEGF. Ratios of cyclin D1 and GAPDH products are shown ( n =8). ( D-F ) Quantifications of U0126 effects on VEGF-dependent cell proliferation (D) or differentiation (E,F). E5 explants were cultured for 24 hours and BrdU labeled for the last 3 hours ( n =6; * P
    Vegf, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 5755 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    vegf  (Abcam)
    94
    Abcam vegf
    IHC analysis of each tumor tissue collected from the OSCC nude mice model. (n=40) OSCC nude mice were treated by inhibition of <t>HIF-1</t> or NF-κB. A. HIF-1α expression. B. NF-κB expression. C. Ki67 expression. D. <t>VEGF</t> expression (×200).
    Vegf, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 3046 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Regeneron vegf trap
    The effects of <t>VEGF</t> Trap, <t>ranibizumab</t> and bevacizumab on luciferase activation induced by VEGF-A 121 and VEGF-A 165 in HEK293/VEGFR2 cells. a Dose response curves for VEGF-A 121 and VEGF-A 165 with EC 50 values of 70 and 30 pM, respectively. PlGF-2 was not active in this assay. b Serial dilutions of VEGF Trap ( open box ), ranibizumab ( triangle ) or bevacizumab ( closed circle ) were added to HEK293/VEGFR2 cells along with 20 pM of VEGF-A 121 . c Serial dilutions of VEGF Trap ( open box ), ranibizumab ( triangle ) or bevacizumab ( closed circle ) were added to HEK293/VEGFR2 cells along with 20 pM of VEGF-A 165 . The cells were incubated for 6 h and OneGlo luciferase substrate was then added to each well. The plates were read on a luminometer and the data were plotted using a four parameter curve fit with GraphPad Prism. Each point represents a replica of 3 wells at each concentration
    Vegf Trap, supplied by Regeneron, used in various techniques. Bioz Stars score: 92/100, based on 955 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher vegf a
    YAP/TAZ suppress β-catenin– and NICD-mediated DLL4 induction. ( A – C ) YAP/TAZ siRNA sensitized HUVECs to Akt activation in response to 50 ng/mL <t>VEGF-A</t> ( A ) and 400 ng/mL Ang-1 ( B ), and 10% <t>FBS</t> ( C ) for 10 minutes. ( D and E ) DAPT (10 μM, 3 hours) or MK2206 (10 μM, 3 hours) abolished YAP/TAZ siRNA–induced mRNA ( D ) and protein ( E ) expression of DLL4. ( F and G ) DAPT (10 μM, 10 minutes pretreatment) or MK2206 (10 μM, 10 minutes pretreatment) attenuated mRNA ( F ) and protein ( G ) expression of DLL4 induced by Y27632 (10 μM, 3 hours). ( H and I ) β-Catenin siRNA abolished YAP/TAZ siRNA–induced mRNA ( H ) and protein ( I ) expression of DLL4. Data are mean ± SEM of triplicates. ** P
    Vegf A, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1173 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Santa Cruz Biotechnology anti vegf
    Inhibition of <t>VEGF</t> affected the VEGF/p38/MMP-2 signaling pathway in PASMCs from rats with monocrotaline-induced pulmonary arterial hypertension by attenuating the effects of miR-15a-5p inhibition. (A) Expression of VEGF, p-p38, MMP-2, Bax and <t>Bcl-2</t> proteins detected using western blot analysis. Quantification of (B) VEGF, (C) p-p38, (D) MMP-2, (E) Bax and (F) Bcl-2 protein expression. ## P
    Anti Vegf, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1839 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Abcam anti vascular endothelial growth factor
    Reduced dermokine ( DMKN ) expression retards <t>growth</t> of xenograft tumors and inhibits tumor metastasis in vivo . (A) Xenograft tumor volume in GFP lentivirus ( NC ) and DMKN ‐sh RNA ( KD ) groups. (B) Comparison of tumor volume between NC and KD groups. (C) Expression of angiogenesis‐associated proteins in blank (non‐transfected), NC , and KD groups. VEGF , <t>vascular</t> <t>endothelial</t> growth <t>factor.</t> (D) Expression of angiogenesis‐associated mRNA in NC and KD groups. (E) Immunohistochemical staining for the vascular marker CD 31 in NC and KD xenograft tumor tissue samples. (G) Fluorescence intensity analysis indicated that reduced DMKN levels inhibit pancreatic cancer metastasis in vivo .
    Anti Vascular Endothelial Growth Factor, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    R&D Systems vegf
    Pleiotropic effects of <t>NGF</t> on HUVEC . (A) Growth assay. HUVEC cultured on standard culture plastic were treated with NGF (100 ng/ml), or <t>VEGF</t> (10 ng/ml) for 48 h. (B and C) Migration and invasion assay using Transwells. (D) Cord formation assay on matrigel-coated 24-well dishes. (E) Endothelial cell monolayer permeability assay. The kinetics of 70 kDa Dextran-FITC having passed through the monolayer of endothelial cells was determined by measurement of fluorescence in the lower chambers at different time points. A to D Student's t-test, *p
    Vegf, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 4694 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    R&D Systems human vegf
    Inhibition of <t>VEGF</t> and <t>NGF</t> by specific neutralizing antibodies significantly suppressed the effects of hDPSC transplantation on the MNCV ( a ) and SNCV ( b ). The neutralizing antibodies against VEGF and NGF were continuously administered using an osmotic pump. The pump was inserted on the day of hDPSC transplantation. The results are expressed as the mean ± SD ( n = 6). ** P
    Human Vegf, supplied by R&D Systems, used in various techniques. Bioz Stars score: 99/100, based on 867 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    R&D Systems recombinant human vegf a
    <t>VEGF</t> pellet implantation induces blood and lymphatic vessel formation In vivo observation of angiogenesis and lymphangiogenesis in a Prox1-GFP/Flk1::myr-mCherry mouse over 10 days following 150ng VEGF pellet implantation (left 3 columns) or control PBS pellet implantation (far right column). ( A-P ) SteREO Lumar microscopy images of GFP-expressing lymphatic vessels (green), mCherry-expressing blood vessels (red) and overlays (right two columns): ( A-D ) prior to implantation, ( E-H ) 3 days post-implantation, ( I-L ) 7 days post-implantation, and ( MP ) 10 days post-implantation. ( Q-T ) Confocal imaging of the same corneas on day 10. Scale bar: 500 μm in ( A-P ) and 200 μm in ( Q-T ). Arrows ( A ): regularly spaced lymphatic vessels penetrating the cornea in the uninjured eye; arrowheads ( E ): new lymphatic vessels budding from the cornea 3 days after VEGF pellet implantation; asterisk ( I ): one potential new lymphatic vessel budding from the cornea on day 7 after the initial phase; circle ( T ): outline of the implanted PBS control pellet.
    Recombinant Human Vegf A, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 279 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore vascular endothelial growth factor vegf
    Effect of high calorie environment towards oxidative stress and pro-inflammatory status in rat strains. After the completion of feeding experiment, blood was collected; plasma was separated and estimated the levels of pro-inflammatory cytokines. ( A ) Interleukin 6; IL-6. ( B ) Tumor necrosis factor alpha; TNFα. ( C ) Interleukin 1 beta; IL-1β. ( D ) Macrophage inflammatory protein 1 alpha; MIP-1α. ( E ) Vascular endothelial growth factor; <t>VEGF.</t> ( F ) <t>IFN</t> gamma inducible protein 10; IP-10. ( G ) Monocyte chemotactic protein 1; MCP1 and anti -inflammatory cytokines such as ( H ) Interleukin 4; IL-4 and ( I ) Interleukin 10; IL-10. ( J ) TBARS assay was performed on liver tissue lysate to study oxidative stress. Data was presented as mean ± SEM (n = 6 per group). Diets: control; HF, high fat; HS, high sucrose; HFS, high fat sucrose. TBARS, Thio Barbituricacid Reactive Substances. *P
    Vascular Endothelial Growth Factor Vegf, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 513 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    PeproTech vegf
    pEVs inhibit angiogenesis in vivo . (a) 12-week-old male C57BL/6J mice were subcutaneously injected with either 5 µg of cEVs and pEVs, mixed with <t>Matrigel</t> Matrix supplemented with <t>VEGF</t> 100 ng/mL (450-32 Peprotech) and Heparin 50 units/mL. After 7 days, plugs were harvested. For haemoglobin quantification, plugs were processed by TissueLyser and the haemoglobin content was measured using Drabkin’s reagent kit 525 (Sigma-Aldrich). Each value was first normalized on the total plug protein quantity, measured by BCA assay, and then on the negative control (plugs with vehicle). (b, c) 1-day-old C57BL/6J mouse pups were intraperitoneally injected with a total of 10 μg of cEVs or pEVs, using PBS as control. Mice were sacrificed for retina collection. Dissected retina were stained with isolectin-b4 (green) and digital images were captured using inverted fluorescence confocal microscope. Analyses of the relative radial expansion and of the relative branching point were performed. All data are expressed as means ± SEM, normalized on control (mice treated with vehicle) (n = 8 mice/group). Ordinary one-way ANOVA; * P
    Vegf, supplied by PeproTech, used in various techniques. Bioz Stars score: 99/100, based on 1475 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    R&D Systems recombinant human vegf165
    One-day post–spinal cord injury (SCI) samples preincubated with vascular endothelial growth factor (VEGF) antibody. ( A ) Western analysis of 24-h samples indicating a reduction in the <t>VEGF165</t> isoform. ( B ) A Western blot of VEGF165 samples at 2
    Recombinant Human Vegf165, supplied by R&D Systems, used in various techniques. Bioz Stars score: 98/100, based on 427 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Cell Signaling Technology Inc vegf
    Effect of miR-126-5p on HUVEC proliferation and downstream signaling pathways. ( A ) qRT-PCR results showing the expression of miR-126-5p in the different groups. ( B ) Representative western blot showing the expression of p-Akt, <t>VEGF,</t> eNOS and <t>CD31</t> in each group (normalized to the expression of β-tubulin). ( C ) Densitometry analyses of p-Akt, VEGF, eNOS and CD31 expression in each group normalized to the expression of t-Akt and β-tubulin. The error bars represent the ±SDs. *P
    Vegf, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 684 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    R&D Systems mouse vegf r3 flt 4 antibody
    Effect of miR-126-5p on HUVEC proliferation and downstream signaling pathways. ( A ) qRT-PCR results showing the expression of miR-126-5p in the different groups. ( B ) Representative western blot showing the expression of p-Akt, <t>VEGF,</t> eNOS and <t>CD31</t> in each group (normalized to the expression of β-tubulin). ( C ) Densitometry analyses of p-Akt, VEGF, eNOS and CD31 expression in each group normalized to the expression of t-Akt and β-tubulin. The error bars represent the ±SDs. *P
    Mouse Vegf R3 Flt 4 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 452 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Santa Cruz Biotechnology rabbit anti vegf
    Endogenous neurogenesis was increased in the NSC-transplanted Tg2576 mice. ( a ) At 2.5 months after transplantation, endogenous neurogenesis was observed in the DG of hippocampus by immunohistochemisty using anti-DCX antibody. DCX-positive cells were calculated in mm 2 area of DG. Graph represents that endogenous neurogenesis was enhanced by NSC transplantation. ( b ) The levels of <t>PSA-NCAM</t> were analyzed by immunohistochemistry. In Tg-NSC group, the levels of PSA-NCAM were increased compared with the Tg-sham group. Graph represents that PSA-NCAM was enhanced by NSC transplantation. ( c ) In Tg-NSC group, the levels of PSA-NCAM and <t>VEGF</t> were increased compared with the Tg-sham group. Quantitative analysis shows that NSC transplantation significantly increased the level of PSA-NCAM and VEGF expressions. * P
    Rabbit Anti Vegf, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 531 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    R&D Systems recombinant vegf
    <t>SLPI</t> promotes proliferation of adult neural stem cells and induces cyclin D1 . A: Proliferation of rat neural stem cells after treatment with SLPI or <t>VEGF,</t> respectively. Cells were treated with indicated amounts of SLPI or VEGF for three days. Afterwards, they were pulsed with 10 μM BrdU. Proportion of BrdU-positive cells (+ std dev) was determined with the FITC BrdU Flow Kit. Presentation of a representative result of three experiments. *:p
    Recombinant Vegf, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 254 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Genentech anti vegf antibody
    Long-term effects of intravitreally injected <t>anti-VEGF</t> antibody on BAT. (A) Quantitative analyses of the number of large lipid droplets ( > 50 μm 2 ) per field at x400 magnification ( n = 3–6). The effects of anti-VEGF antibody were quantitatively analyzed by comparison to the group treated with intravitreal <t>PBS</t> injection as 100%. (B) Quantitative analyses of vascularity of interscapular BAT demonstrated by isolectin B4 staining ( n = 3–6). The effects of anti-VEGF antibody were quantitatively analyzed by comparison to the group treated with intravitreal PBS injection as 100%. (C) The changes in body weight from P14 to P56. Anti-VEGF, anti-VEGF antibody. NS , not significant (two-tailed, unpaired T-test).
    Anti Vegf Antibody, supplied by Genentech, used in various techniques. Bioz Stars score: 92/100, based on 118 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    R&D Systems anti vegf antibody
    Neurotrophic factor expression in mesenchymal stem cells. ( A ) RT-PCR analysis of neurotrophic factor transcripts NGF, GDNF, NT-3, <t>BDNF,</t> <t>VEGF</t> from undifferentiated MSC (uMSC) and differentiated MSC (dMSC) harvested from young and old donors. Amplicon size is shown in base pairs (bp). ( B and C ) ELISA was used to determine BDNF and VEGF protein levels in cell culture supernatants. ***P
    Anti Vegf Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 185 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    VASH1 inhibits angiogenesis and lymphangiogenesis induced by VEGF-A. A: Pellets containing 160 ng of VEGF-A plus or minus 4 ng of VASH1 were inoculated into the mouse cornea. Fourteen days after the inoculation, the corneas were harvested and immunostained

    Journal: The American Journal of Pathology

    Article Title: Endogenous Angiogenesis Inhibitor Vasohibin1 Exhibits Broad-Spectrum Antilymphangiogenic Activity and Suppresses Lymph Node Metastasis

    doi: 10.2353/ajpath.2010.090829

    Figure Lengend Snippet: VASH1 inhibits angiogenesis and lymphangiogenesis induced by VEGF-A. A: Pellets containing 160 ng of VEGF-A plus or minus 4 ng of VASH1 were inoculated into the mouse cornea. Fourteen days after the inoculation, the corneas were harvested and immunostained

    Article Snippet: Briefly, 4-week-old male BALB/c mice (Charles River Laboratories Japan, Inc., Yokohama, Japan) were deeply anesthetized, and 0.3 μg of poly-2-hydroxyethyl methacrylate (HEME) pellets (Sigma, St. Louis, Mo, USA) containing either vehicle or 160 ng of VEGF-A (VEGF165 , Sigma), 160 ng of VEGF-CCys156ser (R & D Systems, Inc., Minneapolis, MN), 12.5 ng or 80 ng of FGF2 (BD Biosciences, San Jose, CA), or 80 ng of PDGF-BB (R & D Systems, Inc.) was implanted in the corneas.

    Techniques:

    Effects of MEK-ERK inhibition on VEGF-dependent proliferation and differentiation. ( A ) Western blots of VEGF-induced phospho-ERK. E6 retinas were cultured with or without U0126 for 60 minutes before VEGF was added for 10 minutes. Blots were probed with phospho-ERK1/2 (top) and α-tubulin (bottom) antibodies.( B ) Quantification of optical densities of phospho-ERK signals were normalized against α-tubulin signals ( n =4). ( C ) RT-PCR detection of cyclin D1 transcripts in E6 retinas cultured for 24 hours in the presence or absence of VEGF. Ratios of cyclin D1 and GAPDH products are shown ( n =8). ( D-F ) Quantifications of U0126 effects on VEGF-dependent cell proliferation (D) or differentiation (E,F). E5 explants were cultured for 24 hours and BrdU labeled for the last 3 hours ( n =6; * P

    Journal: Development (Cambridge, England)

    Article Title: VEGF activates divergent intracellular signaling components to regulate retinal progenitor cell proliferation and neuronal differentiation

    doi: 10.1242/dev.02385

    Figure Lengend Snippet: Effects of MEK-ERK inhibition on VEGF-dependent proliferation and differentiation. ( A ) Western blots of VEGF-induced phospho-ERK. E6 retinas were cultured with or without U0126 for 60 minutes before VEGF was added for 10 minutes. Blots were probed with phospho-ERK1/2 (top) and α-tubulin (bottom) antibodies.( B ) Quantification of optical densities of phospho-ERK signals were normalized against α-tubulin signals ( n =4). ( C ) RT-PCR detection of cyclin D1 transcripts in E6 retinas cultured for 24 hours in the presence or absence of VEGF. Ratios of cyclin D1 and GAPDH products are shown ( n =8). ( D-F ) Quantifications of U0126 effects on VEGF-dependent cell proliferation (D) or differentiation (E,F). E5 explants were cultured for 24 hours and BrdU labeled for the last 3 hours ( n =6; * P

    Article Snippet: Blots were incubated with antibodies against AP (Zymed), VEGF (SantaCruz), FLAG (Sigma), GFP (Molecular Probe), α-tubulin (Sigma) or phospho-ERK1/2 (Cell Signaling), followed by secondary antibodies conjugated with horseradish peroxidase (HRP) and detected by enhanced chemiluminescence (ECL plus, Amersham).

    Techniques: Inhibition, Western Blot, Cell Culture, Reverse Transcription Polymerase Chain Reaction, Labeling

    Influence of VEGF on retinal cell proliferation. ( A ) Effects of VEGF concentrations on BrdU incorporation in vitro. E5–E7 explants were labeled with BrdU for the last 6 hours ( n =4 or 5). ( B-D ) Western blots show sFLK1 (B), VEGF (C) or FLK1-DN (D) expression. Culture media of transfected HEK cells (B,C) or infected DF-1 cell extracts (D) were probed with antibodies against AP (B), VEGF (C) or FLAG tag (D). Controls used were CMV-AP (B,C) or RCAS-AP virus (D). Arrowheads indicate bands with expected molecular weights. ( E-G ) Immunostaining show effects of AP (E), sFLK1 (F) or VEGF (G) in E5–E7 explants on BrdU labeling for the last 3 hours. Scale bars: 50 μm. ( H ) Effects of sFLK1-and VEGF-producing cells on BrdU incorporation (3 hours)at E5 in vivo. Black and white bars represent implanted and the contralateral non-implanted eyes, respectively ( n =5 or 6; * P

    Journal: Development (Cambridge, England)

    Article Title: VEGF activates divergent intracellular signaling components to regulate retinal progenitor cell proliferation and neuronal differentiation

    doi: 10.1242/dev.02385

    Figure Lengend Snippet: Influence of VEGF on retinal cell proliferation. ( A ) Effects of VEGF concentrations on BrdU incorporation in vitro. E5–E7 explants were labeled with BrdU for the last 6 hours ( n =4 or 5). ( B-D ) Western blots show sFLK1 (B), VEGF (C) or FLK1-DN (D) expression. Culture media of transfected HEK cells (B,C) or infected DF-1 cell extracts (D) were probed with antibodies against AP (B), VEGF (C) or FLAG tag (D). Controls used were CMV-AP (B,C) or RCAS-AP virus (D). Arrowheads indicate bands with expected molecular weights. ( E-G ) Immunostaining show effects of AP (E), sFLK1 (F) or VEGF (G) in E5–E7 explants on BrdU labeling for the last 3 hours. Scale bars: 50 μm. ( H ) Effects of sFLK1-and VEGF-producing cells on BrdU incorporation (3 hours)at E5 in vivo. Black and white bars represent implanted and the contralateral non-implanted eyes, respectively ( n =5 or 6; * P

    Article Snippet: Blots were incubated with antibodies against AP (Zymed), VEGF (SantaCruz), FLAG (Sigma), GFP (Molecular Probe), α-tubulin (Sigma) or phospho-ERK1/2 (Cell Signaling), followed by secondary antibodies conjugated with horseradish peroxidase (HRP) and detected by enhanced chemiluminescence (ECL plus, Amersham).

    Techniques: BrdU Incorporation Assay, In Vitro, Labeling, Western Blot, Expressing, Transfection, Infection, FLAG-tag, Immunostaining, In Vivo

    Requirements of HES1 in VEGF-dependent retinal proliferation and differentiation. E5 retinas were co-electroporated with dnHES1- and GFP-expressing constructs, and then cultured as explants ( A , B ) or dissociated cells in collagen gels ( C , D ). VEGF was added at 24 hours post transfection and BrdU was added for the last 3 hours. (A,B) Effects of dnHES1 on Brn3a + cells at E8 (A) or BrdU + cells at E7 (B) among transfected GFP + cells ( n =6; * P

    Journal: Development (Cambridge, England)

    Article Title: VEGF activates divergent intracellular signaling components to regulate retinal progenitor cell proliferation and neuronal differentiation

    doi: 10.1242/dev.02385

    Figure Lengend Snippet: Requirements of HES1 in VEGF-dependent retinal proliferation and differentiation. E5 retinas were co-electroporated with dnHES1- and GFP-expressing constructs, and then cultured as explants ( A , B ) or dissociated cells in collagen gels ( C , D ). VEGF was added at 24 hours post transfection and BrdU was added for the last 3 hours. (A,B) Effects of dnHES1 on Brn3a + cells at E8 (A) or BrdU + cells at E7 (B) among transfected GFP + cells ( n =6; * P

    Article Snippet: Blots were incubated with antibodies against AP (Zymed), VEGF (SantaCruz), FLAG (Sigma), GFP (Molecular Probe), α-tubulin (Sigma) or phospho-ERK1/2 (Cell Signaling), followed by secondary antibodies conjugated with horseradish peroxidase (HRP) and detected by enhanced chemiluminescence (ECL plus, Amersham).

    Techniques: Expressing, Construct, Cell Culture, Transfection

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

    Journal: PLoS ONE

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

    doi: 10.1371/journal.pone.0023914

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

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

    Techniques:

    IHC analysis of each tumor tissue collected from the OSCC nude mice model. (n=40) OSCC nude mice were treated by inhibition of HIF-1 or NF-κB. A. HIF-1α expression. B. NF-κB expression. C. Ki67 expression. D. VEGF expression (×200).

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Co-expression of HIF-1 and TLR3 is associated with poor prognosis in oral squamous cell carcinoma

    doi:

    Figure Lengend Snippet: IHC analysis of each tumor tissue collected from the OSCC nude mice model. (n=40) OSCC nude mice were treated by inhibition of HIF-1 or NF-κB. A. HIF-1α expression. B. NF-κB expression. C. Ki67 expression. D. VEGF expression (×200).

    Article Snippet: DAKO was used with HIF-1α antibody (1:200, Abcam), TLR3 (1:100, Abcam), Ki67 (1:300, Abcam), VEGF (1:250, Abcam), NF-κB p65 (1:150, Cell Signaling Technology).

    Techniques: Immunohistochemistry, Mouse Assay, Inhibition, Expressing

    Proposed mechanisms underlying the therapeutic actions of TRPC1 knockdown via Lipofectamine delivered siRNA against hypoxia-induced pulmonary arterial hypertension (PAH) in a murine model of pulmonary arterial hypertension (PAH). α -SMA: alpha-smooth muscle actin; BMP-2: bone morphogenetic protein-2; c-Csp3: cleaved caspase-3; c-PARP: cleaved poly(ADP-ribose) polymerase; eNOS: endothelial nitric oxide synthase; ET-1: endothelin-1; HIF-1 α : hypoxia-inducible factor 1-alpha; HW: heart weight; MHC: myosin heavy chain; mito-Bax: mitochondrial Bax; MMP-9: matrix metalloproteinase-9; p-Smad: phosphorylated Smad; RVSP: right ventricle systolic pressure; TGF- β : transforming growth factor beta; TNF- α : tumor necrosis factor alpha; TRPC: transient receptor potential cation channel; VEGF: vascular endothelial growth factor.

    Journal: Stem Cells International

    Article Title: Reducing TRPC1 Expression through Liposome-Mediated siRNA Delivery Markedly Attenuates Hypoxia-Induced Pulmonary Arterial Hypertension in a Murine Model

    doi: 10.1155/2014/316214

    Figure Lengend Snippet: Proposed mechanisms underlying the therapeutic actions of TRPC1 knockdown via Lipofectamine delivered siRNA against hypoxia-induced pulmonary arterial hypertension (PAH) in a murine model of pulmonary arterial hypertension (PAH). α -SMA: alpha-smooth muscle actin; BMP-2: bone morphogenetic protein-2; c-Csp3: cleaved caspase-3; c-PARP: cleaved poly(ADP-ribose) polymerase; eNOS: endothelial nitric oxide synthase; ET-1: endothelin-1; HIF-1 α : hypoxia-inducible factor 1-alpha; HW: heart weight; MHC: myosin heavy chain; mito-Bax: mitochondrial Bax; MMP-9: matrix metalloproteinase-9; p-Smad: phosphorylated Smad; RVSP: right ventricle systolic pressure; TGF- β : transforming growth factor beta; TNF- α : tumor necrosis factor alpha; TRPC: transient receptor potential cation channel; VEGF: vascular endothelial growth factor.

    Article Snippet: The membranes were incubated with the indicated primary antibodies [TRPC1 (1 : 1500, Abcam), TRPC4 (1 : 600, Abcam), TRPC6 (1 : 1500, Abcam), Bax (1 : 1000, Abcam), caspase-3 (1 : 1000, Cell Signaling), poly(ADP-ribose) polymerase (PARP) (1 : 1000, Cell Signaling), protein expressions of Bcl-2 (1 : 200, Abcam), phosphorylated (p)-Smad3 (1 : 1000, Cell Signaling), p-Smad1/5 (1 : 1000, Cell Signaling), bone morphogenetic protein- (BMP-) 2 (1 : 5000, Abcam), transforming growth factor (TGF)-β (1 : 500, Abcam), hypoxia-induced factor- (HIF-) 1α (1 : 750, Abcam), vascular endothelial growth factor (VEGF) (1 : 1000, Abcam), Ku-70 (1 : 1000, Abcam), and actin (1 : 10000, Chemicon)] for 1 hour at room temperature.

    Techniques:

    Protein expressions of TRPCs, HIF-1 α , and VEGF of lung parenchyma by day 28 after hypoxia-induced pulmonary arterial hypertension (PAH) ( n = 10). (a) Protein expression of TRPC1, ∗ versus other groups with different symbols (∗, †, ‡), P

    Journal: Stem Cells International

    Article Title: Reducing TRPC1 Expression through Liposome-Mediated siRNA Delivery Markedly Attenuates Hypoxia-Induced Pulmonary Arterial Hypertension in a Murine Model

    doi: 10.1155/2014/316214

    Figure Lengend Snippet: Protein expressions of TRPCs, HIF-1 α , and VEGF of lung parenchyma by day 28 after hypoxia-induced pulmonary arterial hypertension (PAH) ( n = 10). (a) Protein expression of TRPC1, ∗ versus other groups with different symbols (∗, †, ‡), P

    Article Snippet: The membranes were incubated with the indicated primary antibodies [TRPC1 (1 : 1500, Abcam), TRPC4 (1 : 600, Abcam), TRPC6 (1 : 1500, Abcam), Bax (1 : 1000, Abcam), caspase-3 (1 : 1000, Cell Signaling), poly(ADP-ribose) polymerase (PARP) (1 : 1000, Cell Signaling), protein expressions of Bcl-2 (1 : 200, Abcam), phosphorylated (p)-Smad3 (1 : 1000, Cell Signaling), p-Smad1/5 (1 : 1000, Cell Signaling), bone morphogenetic protein- (BMP-) 2 (1 : 5000, Abcam), transforming growth factor (TGF)-β (1 : 500, Abcam), hypoxia-induced factor- (HIF-) 1α (1 : 750, Abcam), vascular endothelial growth factor (VEGF) (1 : 1000, Abcam), Ku-70 (1 : 1000, Abcam), and actin (1 : 10000, Chemicon)] for 1 hour at room temperature.

    Techniques: Expressing

    The effects of VEGF Trap, ranibizumab and bevacizumab on luciferase activation induced by VEGF-A 121 and VEGF-A 165 in HEK293/VEGFR2 cells. a Dose response curves for VEGF-A 121 and VEGF-A 165 with EC 50 values of 70 and 30 pM, respectively. PlGF-2 was not active in this assay. b Serial dilutions of VEGF Trap ( open box ), ranibizumab ( triangle ) or bevacizumab ( closed circle ) were added to HEK293/VEGFR2 cells along with 20 pM of VEGF-A 121 . c Serial dilutions of VEGF Trap ( open box ), ranibizumab ( triangle ) or bevacizumab ( closed circle ) were added to HEK293/VEGFR2 cells along with 20 pM of VEGF-A 165 . The cells were incubated for 6 h and OneGlo luciferase substrate was then added to each well. The plates were read on a luminometer and the data were plotted using a four parameter curve fit with GraphPad Prism. Each point represents a replica of 3 wells at each concentration

    Journal: Angiogenesis

    Article Title: Binding and neutralization of vascular endothelial growth factor (VEGF) and related ligands by VEGF Trap, ranibizumab and bevacizumab

    doi: 10.1007/s10456-011-9249-6

    Figure Lengend Snippet: The effects of VEGF Trap, ranibizumab and bevacizumab on luciferase activation induced by VEGF-A 121 and VEGF-A 165 in HEK293/VEGFR2 cells. a Dose response curves for VEGF-A 121 and VEGF-A 165 with EC 50 values of 70 and 30 pM, respectively. PlGF-2 was not active in this assay. b Serial dilutions of VEGF Trap ( open box ), ranibizumab ( triangle ) or bevacizumab ( closed circle ) were added to HEK293/VEGFR2 cells along with 20 pM of VEGF-A 121 . c Serial dilutions of VEGF Trap ( open box ), ranibizumab ( triangle ) or bevacizumab ( closed circle ) were added to HEK293/VEGFR2 cells along with 20 pM of VEGF-A 165 . The cells were incubated for 6 h and OneGlo luciferase substrate was then added to each well. The plates were read on a luminometer and the data were plotted using a four parameter curve fit with GraphPad Prism. Each point represents a replica of 3 wells at each concentration

    Article Snippet: The results of these experiments show that VEGF Trap binds to VEGF-A with higher affinity and a faster association rate than ranibizumab or bevacizumab, and that VEGF Trap has the unique ability to additionally bind VEGF-B and PlGF.

    Techniques: Luciferase, Activation Assay, Incubation, Concentration Assay

    The effects of VEGF Trap, ranibizumab and bevacizumab on HUVEC migration. a HUVEC were placed in the upper compartment of the Boyden chamber and allowed to migrate towards basal media containing 0.1% fetal bovine serum with or without VEGF-A 165 or VEGF-A 165 mixed with four concentrations each of VEGF Trap ( circles , solid line ), ranibizumab ( triangles , dotted line ) or bevacizumab ( squares , dashed line ) ranging from 0.013 to 13 nM. The percentage of total migration ( y -axis) was calculated as ( F Drug − F Basal )/( F Total − F Basal ) × 100; where F Total is fluorescence in the presence of VEGF-A 165 , F Basal is fluorescence in the absence of VEGF-A 165 , and F Drug is fluorescence in the presence of VEGF-A 165 mixed with drug at a specific molar ratio ( x -axis). b HUVEC migration was assessed in the absence and presence of human PLGF-2 (hPLGF-2) or mouse PLGF-2 (mPLGF-2) with and without a 100-fold molar excess of VEGF Trap (VGT), ranibizumab (RAN) or bevacizumab (BEV). Fold migration ( y -axis) was calculated as the ratio F / F Basal ; where F is the total fluorescence measured for the indicated condition ( x -axis) and F Basal is the fluorescence in the absence of either hPLGF-2 or mPLGF-2. Statistical significance: * P

    Journal: Angiogenesis

    Article Title: Binding and neutralization of vascular endothelial growth factor (VEGF) and related ligands by VEGF Trap, ranibizumab and bevacizumab

    doi: 10.1007/s10456-011-9249-6

    Figure Lengend Snippet: The effects of VEGF Trap, ranibizumab and bevacizumab on HUVEC migration. a HUVEC were placed in the upper compartment of the Boyden chamber and allowed to migrate towards basal media containing 0.1% fetal bovine serum with or without VEGF-A 165 or VEGF-A 165 mixed with four concentrations each of VEGF Trap ( circles , solid line ), ranibizumab ( triangles , dotted line ) or bevacizumab ( squares , dashed line ) ranging from 0.013 to 13 nM. The percentage of total migration ( y -axis) was calculated as ( F Drug − F Basal )/( F Total − F Basal ) × 100; where F Total is fluorescence in the presence of VEGF-A 165 , F Basal is fluorescence in the absence of VEGF-A 165 , and F Drug is fluorescence in the presence of VEGF-A 165 mixed with drug at a specific molar ratio ( x -axis). b HUVEC migration was assessed in the absence and presence of human PLGF-2 (hPLGF-2) or mouse PLGF-2 (mPLGF-2) with and without a 100-fold molar excess of VEGF Trap (VGT), ranibizumab (RAN) or bevacizumab (BEV). Fold migration ( y -axis) was calculated as the ratio F / F Basal ; where F is the total fluorescence measured for the indicated condition ( x -axis) and F Basal is the fluorescence in the absence of either hPLGF-2 or mPLGF-2. Statistical significance: * P

    Article Snippet: The results of these experiments show that VEGF Trap binds to VEGF-A with higher affinity and a faster association rate than ranibizumab or bevacizumab, and that VEGF Trap has the unique ability to additionally bind VEGF-B and PlGF.

    Techniques: Migration, Fluorescence

    The effects of VEGF Trap, ranibizumab and bevacizumab on calcium mobilization induced byVEGF-A 165 in HUVEC. a A dose–response curve generated using serial dilutions of VEGF-A 165 (4.0 nM–0.023 pM) resulted in an EC 50 value of 5 pM. b Serial dilutions of VEGF Trap ( open box ), ranibizumab ( triangle ) or bevacizumab ( closed circle ) were added to HUVEC along with 20 pM of VEGF-A 165 . The VEGF-A 165 was preincubated with the inhibitors for 10 min at 25°C. The solution was added to HUVEC preloaded with fluo-4 and the fluorescence of the well was determined on a FLIPR instrument. The data were plotted using a four parameter curve fit with GraphPad Prism. Each point represents duplicate wells at each concentration

    Journal: Angiogenesis

    Article Title: Binding and neutralization of vascular endothelial growth factor (VEGF) and related ligands by VEGF Trap, ranibizumab and bevacizumab

    doi: 10.1007/s10456-011-9249-6

    Figure Lengend Snippet: The effects of VEGF Trap, ranibizumab and bevacizumab on calcium mobilization induced byVEGF-A 165 in HUVEC. a A dose–response curve generated using serial dilutions of VEGF-A 165 (4.0 nM–0.023 pM) resulted in an EC 50 value of 5 pM. b Serial dilutions of VEGF Trap ( open box ), ranibizumab ( triangle ) or bevacizumab ( closed circle ) were added to HUVEC along with 20 pM of VEGF-A 165 . The VEGF-A 165 was preincubated with the inhibitors for 10 min at 25°C. The solution was added to HUVEC preloaded with fluo-4 and the fluorescence of the well was determined on a FLIPR instrument. The data were plotted using a four parameter curve fit with GraphPad Prism. Each point represents duplicate wells at each concentration

    Article Snippet: The results of these experiments show that VEGF Trap binds to VEGF-A with higher affinity and a faster association rate than ranibizumab or bevacizumab, and that VEGF Trap has the unique ability to additionally bind VEGF-B and PlGF.

    Techniques: Generated, Fluorescence, Concentration Assay

    The effects of VEGF Trap, ranibizumab and bevacizumab on luciferase activation induced by VEGF-A 121 , VEGF-A 165 , human PlGF-2 (hPlGF-2) or mouse PlGF-2 (mPLGF-2) in HEK293/VEGFR1 cells. a Dose response curves for VEGF-A 121 , VEGF-A 165 and hPlGF-2 yielded EC 50 values of 13, 17, and 29 pM, respectively. b Serial dilutions of VEGF Trap ( open box ), ranibizumab ( triangle ), or bevacizumab ( closed circle ) were added to HEK293/VEGFR1 cells along with 20 pM of VEGF-A 121 . c Serial dilutions of VEGF Trap ( open box ), ranibizumab ( triangle ), or bevacizumab ( closed circle ) were added to HEK293/VEGFR1 cells along with 20 pM of VEGF-A 165 . d Serial dilutions of VEGF Trap ( open box ), ranibizumab ( triangle ), or bevacizumab ( closed circle ) were added to HEK293/VEGFR1 cells along with 40 pM of human PlGF-2. e Dose response curve for mPlGF-2 yielded an EC 50 value of 10 pM ( f ). Serial dilutions of VEGF Trap were added to HEK293/VEGFR1 cells along with 20 pM of mPlGF-2. The cells were incubated for 6 h and OneGlo luciferase substrate was then added to each well. The plates were read on a luminometer and the data were plotted using a four parameter curve fit with GraphPad Prism. Each point represents a replica of 3 wells at each concentration

    Journal: Angiogenesis

    Article Title: Binding and neutralization of vascular endothelial growth factor (VEGF) and related ligands by VEGF Trap, ranibizumab and bevacizumab

    doi: 10.1007/s10456-011-9249-6

    Figure Lengend Snippet: The effects of VEGF Trap, ranibizumab and bevacizumab on luciferase activation induced by VEGF-A 121 , VEGF-A 165 , human PlGF-2 (hPlGF-2) or mouse PlGF-2 (mPLGF-2) in HEK293/VEGFR1 cells. a Dose response curves for VEGF-A 121 , VEGF-A 165 and hPlGF-2 yielded EC 50 values of 13, 17, and 29 pM, respectively. b Serial dilutions of VEGF Trap ( open box ), ranibizumab ( triangle ), or bevacizumab ( closed circle ) were added to HEK293/VEGFR1 cells along with 20 pM of VEGF-A 121 . c Serial dilutions of VEGF Trap ( open box ), ranibizumab ( triangle ), or bevacizumab ( closed circle ) were added to HEK293/VEGFR1 cells along with 20 pM of VEGF-A 165 . d Serial dilutions of VEGF Trap ( open box ), ranibizumab ( triangle ), or bevacizumab ( closed circle ) were added to HEK293/VEGFR1 cells along with 40 pM of human PlGF-2. e Dose response curve for mPlGF-2 yielded an EC 50 value of 10 pM ( f ). Serial dilutions of VEGF Trap were added to HEK293/VEGFR1 cells along with 20 pM of mPlGF-2. The cells were incubated for 6 h and OneGlo luciferase substrate was then added to each well. The plates were read on a luminometer and the data were plotted using a four parameter curve fit with GraphPad Prism. Each point represents a replica of 3 wells at each concentration

    Article Snippet: The results of these experiments show that VEGF Trap binds to VEGF-A with higher affinity and a faster association rate than ranibizumab or bevacizumab, and that VEGF Trap has the unique ability to additionally bind VEGF-B and PlGF.

    Techniques: Luciferase, Activation Assay, Incubation, Concentration Assay

    YAP/TAZ suppress β-catenin– and NICD-mediated DLL4 induction. ( A – C ) YAP/TAZ siRNA sensitized HUVECs to Akt activation in response to 50 ng/mL VEGF-A ( A ) and 400 ng/mL Ang-1 ( B ), and 10% FBS ( C ) for 10 minutes. ( D and E ) DAPT (10 μM, 3 hours) or MK2206 (10 μM, 3 hours) abolished YAP/TAZ siRNA–induced mRNA ( D ) and protein ( E ) expression of DLL4. ( F and G ) DAPT (10 μM, 10 minutes pretreatment) or MK2206 (10 μM, 10 minutes pretreatment) attenuated mRNA ( F ) and protein ( G ) expression of DLL4 induced by Y27632 (10 μM, 3 hours). ( H and I ) β-Catenin siRNA abolished YAP/TAZ siRNA–induced mRNA ( H ) and protein ( I ) expression of DLL4. Data are mean ± SEM of triplicates. ** P

    Journal: The Journal of Clinical Investigation

    Article Title: Lysophosphatidic acid–induced YAP/TAZ activation promotes developmental angiogenesis by repressing Notch ligand Dll4

    doi: 10.1172/JCI121955

    Figure Lengend Snippet: YAP/TAZ suppress β-catenin– and NICD-mediated DLL4 induction. ( A – C ) YAP/TAZ siRNA sensitized HUVECs to Akt activation in response to 50 ng/mL VEGF-A ( A ) and 400 ng/mL Ang-1 ( B ), and 10% FBS ( C ) for 10 minutes. ( D and E ) DAPT (10 μM, 3 hours) or MK2206 (10 μM, 3 hours) abolished YAP/TAZ siRNA–induced mRNA ( D ) and protein ( E ) expression of DLL4. ( F and G ) DAPT (10 μM, 10 minutes pretreatment) or MK2206 (10 μM, 10 minutes pretreatment) attenuated mRNA ( F ) and protein ( G ) expression of DLL4 induced by Y27632 (10 μM, 3 hours). ( H and I ) β-Catenin siRNA abolished YAP/TAZ siRNA–induced mRNA ( H ) and protein ( I ) expression of DLL4. Data are mean ± SEM of triplicates. ** P

    Article Snippet: After 24 hours, the beads were washed with EGM-2 BulletKit 3 times and suspended in EGM-2 BulletKit containing 2% FBS, 2 mg/mL fibrinogen (Sigma-Aldrich), 0.15 U/mL aprotinin (Sigma-Aldrich), and 10 ng/mL VEGF-A (Invitrogen).

    Techniques: Activation Assay, Expressing

    Early passage ASCs overexpress the neurotropic protein HGF but not VEGF nor the proinflammatory cytokine IL-1 β . ASCs at passage 3 that were cultured in serum-free conditions for 48 hours were compared to the control group of ASCs at passage 5. Both cells and medium were collected and analyzed at mRNA level and at protein level by qRT-PCR and by ELISA, respectively. (a) qRT-PCR analysis of HGF, VEGF, and IL-1 β . (b) ELISA for HGF protein levels. Each experiment was performed a minimum of 3 samples from 3 different patients. Each experiment was performed a minimum of 3 times.

    Journal: Stem Cells International

    Article Title: Adipose-Derived Mesenchymal Stem Cells Migrate and Rescue RPE in the Setting of Oxidative Stress

    doi: 10.1155/2018/9682856

    Figure Lengend Snippet: Early passage ASCs overexpress the neurotropic protein HGF but not VEGF nor the proinflammatory cytokine IL-1 β . ASCs at passage 3 that were cultured in serum-free conditions for 48 hours were compared to the control group of ASCs at passage 5. Both cells and medium were collected and analyzed at mRNA level and at protein level by qRT-PCR and by ELISA, respectively. (a) qRT-PCR analysis of HGF, VEGF, and IL-1 β . (b) ELISA for HGF protein levels. Each experiment was performed a minimum of 3 samples from 3 different patients. Each experiment was performed a minimum of 3 times.

    Article Snippet: The mRNA expression levels of the growth factors, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-1β (IL-1β ), stromal-derived factor-1 (SDF-1), the chemokine receptor CXCR4, and normalizing housekeeping genes GUSB and RLP27 (see for sequence information) were measured by real-time reverse transcription polymerase chain reaction (RT-PCR) (StepOnePlus, Applied Biosystems) using SYBR® Green qPCR Mastermix (Qiagen).

    Techniques: Cell Culture, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    Inhibition of VEGF affected the VEGF/p38/MMP-2 signaling pathway in PASMCs from rats with monocrotaline-induced pulmonary arterial hypertension by attenuating the effects of miR-15a-5p inhibition. (A) Expression of VEGF, p-p38, MMP-2, Bax and Bcl-2 proteins detected using western blot analysis. Quantification of (B) VEGF, (C) p-p38, (D) MMP-2, (E) Bax and (F) Bcl-2 protein expression. ## P

    Journal: International Journal of Molecular Medicine

    Article Title: MicroRNA-15a-5p induces pulmonary artery smooth muscle cell apoptosis in a pulmonary arterial hypertension model via the VEGF/p38/MMP-2 signaling pathway

    doi: 10.3892/ijmm.2019.4434

    Figure Lengend Snippet: Inhibition of VEGF affected the VEGF/p38/MMP-2 signaling pathway in PASMCs from rats with monocrotaline-induced pulmonary arterial hypertension by attenuating the effects of miR-15a-5p inhibition. (A) Expression of VEGF, p-p38, MMP-2, Bax and Bcl-2 proteins detected using western blot analysis. Quantification of (B) VEGF, (C) p-p38, (D) MMP-2, (E) Bax and (F) Bcl-2 protein expression. ## P

    Article Snippet: The membranes were blocked with 5% skimmed milk in 1X Tris-buffered saline with 0.1% Tween-20 (TBST) followed by incubation overnight at 4°C with the following primary antibodies: Anti-B-cell lymphoma 2 (Bcl-2; cat. no. sc-509; 1:1,000, Santa Cruz Biotechnology, Inc.), anti-Bcl-2-associated X protein (Bax; cat. no. sc-20067; 1:1,000, Santa Cruz Biotechnology, Inc.), anti-VEGF (cat. no. sc-81670; 1:500, Santa Cruz Biotechnology, Inc.), anti-phosphorylated (p)-p38 MAPK (cat. no. sc-17852-R; 1:1,000, Santa Cruz Biotechnology, Inc.), anti-p38 MAPK (cat. no. 8690S; 1:1,000, Cell Signaling Technology, Inc.), anti-MMP-2 (cat. no. sc-53630; 1:1,000, Santa Cruz Biotechnology, Inc.) and anti-GAPDH (cat. no. sc-51631; 1:5,000, Santa Cruz Biotechnology, Inc.).

    Techniques: Inhibition, Expressing, Western Blot

    Upregulation of VEGF affected the VEGF/p38/MMP-2 signaling pathway in PASMCs from rats with monocrotaline-induced pulmonary arterial hypertension by the effects of miR-15a-5p. (A) Results of the protein expression of VEGF, p38, MMP-2, Bax and Bcl-2 measured using western blot analysis. Protein expression of (B) VEGF, (C) p-p38, (D) MMP-2, (E) Bax and (F) Bcl-2. ## P

    Journal: International Journal of Molecular Medicine

    Article Title: MicroRNA-15a-5p induces pulmonary artery smooth muscle cell apoptosis in a pulmonary arterial hypertension model via the VEGF/p38/MMP-2 signaling pathway

    doi: 10.3892/ijmm.2019.4434

    Figure Lengend Snippet: Upregulation of VEGF affected the VEGF/p38/MMP-2 signaling pathway in PASMCs from rats with monocrotaline-induced pulmonary arterial hypertension by the effects of miR-15a-5p. (A) Results of the protein expression of VEGF, p38, MMP-2, Bax and Bcl-2 measured using western blot analysis. Protein expression of (B) VEGF, (C) p-p38, (D) MMP-2, (E) Bax and (F) Bcl-2. ## P

    Article Snippet: The membranes were blocked with 5% skimmed milk in 1X Tris-buffered saline with 0.1% Tween-20 (TBST) followed by incubation overnight at 4°C with the following primary antibodies: Anti-B-cell lymphoma 2 (Bcl-2; cat. no. sc-509; 1:1,000, Santa Cruz Biotechnology, Inc.), anti-Bcl-2-associated X protein (Bax; cat. no. sc-20067; 1:1,000, Santa Cruz Biotechnology, Inc.), anti-VEGF (cat. no. sc-81670; 1:500, Santa Cruz Biotechnology, Inc.), anti-phosphorylated (p)-p38 MAPK (cat. no. sc-17852-R; 1:1,000, Santa Cruz Biotechnology, Inc.), anti-p38 MAPK (cat. no. 8690S; 1:1,000, Cell Signaling Technology, Inc.), anti-MMP-2 (cat. no. sc-53630; 1:1,000, Santa Cruz Biotechnology, Inc.) and anti-GAPDH (cat. no. sc-51631; 1:5,000, Santa Cruz Biotechnology, Inc.).

    Techniques: Expressing, Western Blot

    Western blot analysis of protein expression of IL-6, p-STAT3, survivin, STAT3, and VEGF. Protein levels of IL-6, Survivin, p-STAT3, STAT3, and VEGF in normal gastric and tumor tissue were determined using western blotting. Beta-actin was a loading control. Relative protein expression of IL-6 (A), VEGF (B), surviving (C), p-STAT3 (D) was normalized to of the corresponding beta-actin level. Positive immunoreactive bands were quantified densitometrically and expressed as IL-6, Survivin, p-STAT3, STAT3, and VEGF in optical density units, respectively. * P

    Journal: PLoS ONE

    Article Title: Activation of STAT3 in Human Gastric Cancer Cells via Interleukin (IL)-6-Type Cytokine Signaling Correlates with Clinical Implications

    doi: 10.1371/journal.pone.0075788

    Figure Lengend Snippet: Western blot analysis of protein expression of IL-6, p-STAT3, survivin, STAT3, and VEGF. Protein levels of IL-6, Survivin, p-STAT3, STAT3, and VEGF in normal gastric and tumor tissue were determined using western blotting. Beta-actin was a loading control. Relative protein expression of IL-6 (A), VEGF (B), surviving (C), p-STAT3 (D) was normalized to of the corresponding beta-actin level. Positive immunoreactive bands were quantified densitometrically and expressed as IL-6, Survivin, p-STAT3, STAT3, and VEGF in optical density units, respectively. * P

    Article Snippet: The levels of VEGF, survivin, and IL-6 in the gastric samples were determined by an anti-VEGF antibody (1:100 dilution; Santa Cruz Biotechnology), an anti-survivin antibody (1:100 dilution; Santa Cruz Biotechnology), and an anti-IL-6 antibody (1:50 dilution; Santa Cruz Biotechnology), respectively.

    Techniques: Western Blot, Expressing

    Expression of STAT3 in human gastric cancer tissues. The expression and localization of STAT3, IL-6, VEGF, and survivin in gastric cancer cells were determined using immunohistochemical staining. There was weak or negative expression of STAT3 in adjacent normal mucosa. However, there was strong expression of phosphorylated STAT3 in gastric cancer tissues. The STAT3 staining was mainly localized in the nuclei of tumor epithelial cells, which was indicated by numerous yellowish granules. STAT3 overexpression was associated with with increased expression of IL-6, surviving, and VEGF as well as with increased vessel density (Original magnification of A1-A3 and B1-B3, ×400; A4 and B4, ×200)..

    Journal: PLoS ONE

    Article Title: Activation of STAT3 in Human Gastric Cancer Cells via Interleukin (IL)-6-Type Cytokine Signaling Correlates with Clinical Implications

    doi: 10.1371/journal.pone.0075788

    Figure Lengend Snippet: Expression of STAT3 in human gastric cancer tissues. The expression and localization of STAT3, IL-6, VEGF, and survivin in gastric cancer cells were determined using immunohistochemical staining. There was weak or negative expression of STAT3 in adjacent normal mucosa. However, there was strong expression of phosphorylated STAT3 in gastric cancer tissues. The STAT3 staining was mainly localized in the nuclei of tumor epithelial cells, which was indicated by numerous yellowish granules. STAT3 overexpression was associated with with increased expression of IL-6, surviving, and VEGF as well as with increased vessel density (Original magnification of A1-A3 and B1-B3, ×400; A4 and B4, ×200)..

    Article Snippet: The levels of VEGF, survivin, and IL-6 in the gastric samples were determined by an anti-VEGF antibody (1:100 dilution; Santa Cruz Biotechnology), an anti-survivin antibody (1:100 dilution; Santa Cruz Biotechnology), and an anti-IL-6 antibody (1:50 dilution; Santa Cruz Biotechnology), respectively.

    Techniques: Expressing, Immunohistochemistry, Staining, Over Expression

    Representative Western blot gel and summarized band densities demonstrating protein expression levels of VEGF, VEGF-R1, and VEGF-R2 in P1 aortas. N = 5 per group (each N is pooled protein from 4 different animals from different litters). Error bars indicate standard errors. Asterisks indicate P

    Journal: Reproductive Sciences

    Article Title: Excess Maternal Glucocorticoids in Response to In Utero Undernutrition Inhibit Offspring Angiogenesis

    doi: 10.1177/1933719113508819

    Figure Lengend Snippet: Representative Western blot gel and summarized band densities demonstrating protein expression levels of VEGF, VEGF-R1, and VEGF-R2 in P1 aortas. N = 5 per group (each N is pooled protein from 4 different animals from different litters). Error bars indicate standard errors. Asterisks indicate P

    Article Snippet: For each homogenate, 40 µg of protein (pooled from 4 vessels from 4 animals from different litters) were separated on a 7.5% polyacrylamide gel, then transferred electrophoretically to Immobilon-P membranes (Millipore, Billerica, Massachusetts), blocked for 2 hours in 5% milk, and incubated overnight with antibodies against VEGF (sc-7269), VEGF-R1 (sc-316), MMP-2 (sc-10736), or MMP-9 (sc-10737; Santa Cruz Biotechnology) or antibodies against VEGF-R2 (9698S; Cell Signaling Technology, Beverly, Massachusetts).

    Techniques: Western Blot, Expressing

    Reduced dermokine ( DMKN ) expression retards growth of xenograft tumors and inhibits tumor metastasis in vivo . (A) Xenograft tumor volume in GFP lentivirus ( NC ) and DMKN ‐sh RNA ( KD ) groups. (B) Comparison of tumor volume between NC and KD groups. (C) Expression of angiogenesis‐associated proteins in blank (non‐transfected), NC , and KD groups. VEGF , vascular endothelial growth factor. (D) Expression of angiogenesis‐associated mRNA in NC and KD groups. (E) Immunohistochemical staining for the vascular marker CD 31 in NC and KD xenograft tumor tissue samples. (G) Fluorescence intensity analysis indicated that reduced DMKN levels inhibit pancreatic cancer metastasis in vivo .

    Journal: Cancer Science

    Article Title: Dermokine contributes to epithelial–mesenchymal transition through increased activation of signal transducer and activator of transcription 3 in pancreatic cancer

    doi: 10.1111/cas.13347

    Figure Lengend Snippet: Reduced dermokine ( DMKN ) expression retards growth of xenograft tumors and inhibits tumor metastasis in vivo . (A) Xenograft tumor volume in GFP lentivirus ( NC ) and DMKN ‐sh RNA ( KD ) groups. (B) Comparison of tumor volume between NC and KD groups. (C) Expression of angiogenesis‐associated proteins in blank (non‐transfected), NC , and KD groups. VEGF , vascular endothelial growth factor. (D) Expression of angiogenesis‐associated mRNA in NC and KD groups. (E) Immunohistochemical staining for the vascular marker CD 31 in NC and KD xenograft tumor tissue samples. (G) Fluorescence intensity analysis indicated that reduced DMKN levels inhibit pancreatic cancer metastasis in vivo .

    Article Snippet: The following commercially available antibodies were used: anti‐GAPDH (Bioworld Technology, St. Louis Park, MN, USA), anti‐DMKN (Abcam, Cambridge, MA, USA), anti‐vascular endothelial growth factor (VEGF; Abcam), anti‐ERK1/2, anti‐phospho‐ERK1/2, anti‐AKT, anti‐phospho‐AKT, anti‐STAT3, anti‐phospho‐STAT3, anti‐E‐cadherin, anti‐N‐cadherin, anti‐Snail, anti‐vimentin, anti‐ MMP2, and anti‐MMP9 (all Cell Signaling Technology, Danvers, MA, USA).

    Techniques: Expressing, In Vivo, Transfection, Immunohistochemistry, Staining, Marker, Fluorescence

    Analysis of the oncogenic pathways regulated by dermokine ( DMKN ). (A) Expression of oncogenic pathway proteins in pancreatic ductal adenocarcinoma cells transfected with either DMKN ‐sh RNA ( KD ), GFP lentivirus ( NC ), or non‐transfected (blank) cells. (B) Expression of epithelial–mesenchymal transition ( EMT )‐associated and oncogenic pathway proteins in Patu‐8988 cells treated with U0126 ( MAPK / ERK kinase inhibitor), MK ‐2206 2 HCL ( AKT inhibitor), or STA ‐21 (signal transducer and activator of transcription 3 [ STAT 3] inhibitor). (C) EMT ‐associated proteins and oncogenic pathway proteins in PANC ‐1 cells treated with DMKN overexpressing plasmids. (D) Effect and mechanist model for elucidating the role of DMKN in oncogenic pathway. VEGF , vascular endothelial growth factor.

    Journal: Cancer Science

    Article Title: Dermokine contributes to epithelial–mesenchymal transition through increased activation of signal transducer and activator of transcription 3 in pancreatic cancer

    doi: 10.1111/cas.13347

    Figure Lengend Snippet: Analysis of the oncogenic pathways regulated by dermokine ( DMKN ). (A) Expression of oncogenic pathway proteins in pancreatic ductal adenocarcinoma cells transfected with either DMKN ‐sh RNA ( KD ), GFP lentivirus ( NC ), or non‐transfected (blank) cells. (B) Expression of epithelial–mesenchymal transition ( EMT )‐associated and oncogenic pathway proteins in Patu‐8988 cells treated with U0126 ( MAPK / ERK kinase inhibitor), MK ‐2206 2 HCL ( AKT inhibitor), or STA ‐21 (signal transducer and activator of transcription 3 [ STAT 3] inhibitor). (C) EMT ‐associated proteins and oncogenic pathway proteins in PANC ‐1 cells treated with DMKN overexpressing plasmids. (D) Effect and mechanist model for elucidating the role of DMKN in oncogenic pathway. VEGF , vascular endothelial growth factor.

    Article Snippet: The following commercially available antibodies were used: anti‐GAPDH (Bioworld Technology, St. Louis Park, MN, USA), anti‐DMKN (Abcam, Cambridge, MA, USA), anti‐vascular endothelial growth factor (VEGF; Abcam), anti‐ERK1/2, anti‐phospho‐ERK1/2, anti‐AKT, anti‐phospho‐AKT, anti‐STAT3, anti‐phospho‐STAT3, anti‐E‐cadherin, anti‐N‐cadherin, anti‐Snail, anti‐vimentin, anti‐ MMP2, and anti‐MMP9 (all Cell Signaling Technology, Danvers, MA, USA).

    Techniques: Expressing, Transfection

    Effect of YC-1 on 2-h ischemia-induced VEGF expression in neurons and astrocytes. The cellular distribution of VEGF was analyzed by immunostaining with NeuN (marker of neurons) or GFAP (marker of astrocytes) after 2-h MCAO. Double immunostain of VEGF (green) and NeuN (red) showed a good colocalization of VEGF and neurons and YC-1 treatment significantly decreased the proportion of VEGF-positive neurons (A) . Double immunostain of VEGF (green) and GFAP (red) showed no co-localization of VEGF and astrocytes (B) . After quantification, two-way ANNOVA showed an increased proportion of VEGF positive neurons and YC-1 significantly decreased this upregulation (C,D) . n = 3/group. Scale bar = 50 μm. n = 3/group. * P

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Inhibition of HIF-1α Reduced Blood Brain Barrier Damage by Regulating MMP-2 and VEGF During Acute Cerebral Ischemia

    doi: 10.3389/fncel.2018.00288

    Figure Lengend Snippet: Effect of YC-1 on 2-h ischemia-induced VEGF expression in neurons and astrocytes. The cellular distribution of VEGF was analyzed by immunostaining with NeuN (marker of neurons) or GFAP (marker of astrocytes) after 2-h MCAO. Double immunostain of VEGF (green) and NeuN (red) showed a good colocalization of VEGF and neurons and YC-1 treatment significantly decreased the proportion of VEGF-positive neurons (A) . Double immunostain of VEGF (green) and GFAP (red) showed no co-localization of VEGF and astrocytes (B) . After quantification, two-way ANNOVA showed an increased proportion of VEGF positive neurons and YC-1 significantly decreased this upregulation (C,D) . n = 3/group. Scale bar = 50 μm. n = 3/group. * P

    Article Snippet: Then anti-VEGF antibody (1:200, Abcam), anti-NeuN (1:200, Millipore), anti-GFAP (1:2,000, Millipore) primary antibody were applied to the sections and incubated at 4°C overnight.

    Techniques: Expressing, Immunostaining, Marker

    Pleiotropic effects of NGF on HUVEC . (A) Growth assay. HUVEC cultured on standard culture plastic were treated with NGF (100 ng/ml), or VEGF (10 ng/ml) for 48 h. (B and C) Migration and invasion assay using Transwells. (D) Cord formation assay on matrigel-coated 24-well dishes. (E) Endothelial cell monolayer permeability assay. The kinetics of 70 kDa Dextran-FITC having passed through the monolayer of endothelial cells was determined by measurement of fluorescence in the lower chambers at different time points. A to D Student's t-test, *p

    Journal: Molecular Cancer

    Article Title: Nerve growth factor promotes breast cancer angiogenesis by activating multiple pathways

    doi: 10.1186/1476-4598-9-157

    Figure Lengend Snippet: Pleiotropic effects of NGF on HUVEC . (A) Growth assay. HUVEC cultured on standard culture plastic were treated with NGF (100 ng/ml), or VEGF (10 ng/ml) for 48 h. (B and C) Migration and invasion assay using Transwells. (D) Cord formation assay on matrigel-coated 24-well dishes. (E) Endothelial cell monolayer permeability assay. The kinetics of 70 kDa Dextran-FITC having passed through the monolayer of endothelial cells was determined by measurement of fluorescence in the lower chambers at different time points. A to D Student's t-test, *p

    Article Snippet: Reagents Human recombinant NGF and VEGF, neutralizing antibodies against NGF, VEGF and the corresponding isotype control antibodies were purchased from R & D Systems.

    Techniques: Growth Assay, Cell Culture, Migration, Invasion Assay, Tube Formation Assay, Permeability, Fluorescence

    Involvement of the VEGF in NGF-stimulated angiogenesis . (A) ELISA quantification of VEGF in conditioned media from HUVEC and MDA-MB-231 cells. Cells were treated with NGF (100 ng/ml) for 6 h or 24 h, conditioned media were concentrated before ELISA assay, as described in materials and methods. (B) Invasion assay using Transwells. HUVEC were treated with NGF (100 ng/ml) or VEGF (10 ng/ml) in the presence of isotype control or anti-VEGF neutralizing antibodies (1 μg/ml) for 24 h. (C) In vivo angiogenesis assay. Matrigel containing a mixture of NGF and isotype control or anti-VEGF neutralizing antibodies (37.5 μg/ml) was subcutaneously injected into SCID mice (five mice per group) as described in materials and methods. Hemoglobin in Matrigel plugs was quantified by Drabkin method 7 days after injection. Results are the mean of three independent experiments. Student's t-test, *p

    Journal: Molecular Cancer

    Article Title: Nerve growth factor promotes breast cancer angiogenesis by activating multiple pathways

    doi: 10.1186/1476-4598-9-157

    Figure Lengend Snippet: Involvement of the VEGF in NGF-stimulated angiogenesis . (A) ELISA quantification of VEGF in conditioned media from HUVEC and MDA-MB-231 cells. Cells were treated with NGF (100 ng/ml) for 6 h or 24 h, conditioned media were concentrated before ELISA assay, as described in materials and methods. (B) Invasion assay using Transwells. HUVEC were treated with NGF (100 ng/ml) or VEGF (10 ng/ml) in the presence of isotype control or anti-VEGF neutralizing antibodies (1 μg/ml) for 24 h. (C) In vivo angiogenesis assay. Matrigel containing a mixture of NGF and isotype control or anti-VEGF neutralizing antibodies (37.5 μg/ml) was subcutaneously injected into SCID mice (five mice per group) as described in materials and methods. Hemoglobin in Matrigel plugs was quantified by Drabkin method 7 days after injection. Results are the mean of three independent experiments. Student's t-test, *p

    Article Snippet: Reagents Human recombinant NGF and VEGF, neutralizing antibodies against NGF, VEGF and the corresponding isotype control antibodies were purchased from R & D Systems.

    Techniques: Enzyme-linked Immunosorbent Assay, Multiple Displacement Amplification, Invasion Assay, In Vivo, Angiogenesis Assay, Injection, Mouse Assay

    Angiogenesis assay using Matrigel plugs in SCID mice . Matrigel containing different reagents was subcutaneously injected into SCID mice as described in materials and methods. (A, B) Matrigel was mixed with MDA-MB-231 breast cancer cells and isotype control or anti-NGF neutralizing antibodies (75 μg/ml); (C) Matrigel was mixed with proNGF (7.5 μg/ml), NGF (3.75 μg/ml), or VEGF (0.375 μg/ml). Angiogenesis was analyzed by quantification of hemoglobin (A, C) and microvessel density (B) as described in materials and methods. Five mice were used for each group and results are the mean of three independent experiments. Student's t-test, *p

    Journal: Molecular Cancer

    Article Title: Nerve growth factor promotes breast cancer angiogenesis by activating multiple pathways

    doi: 10.1186/1476-4598-9-157

    Figure Lengend Snippet: Angiogenesis assay using Matrigel plugs in SCID mice . Matrigel containing different reagents was subcutaneously injected into SCID mice as described in materials and methods. (A, B) Matrigel was mixed with MDA-MB-231 breast cancer cells and isotype control or anti-NGF neutralizing antibodies (75 μg/ml); (C) Matrigel was mixed with proNGF (7.5 μg/ml), NGF (3.75 μg/ml), or VEGF (0.375 μg/ml). Angiogenesis was analyzed by quantification of hemoglobin (A, C) and microvessel density (B) as described in materials and methods. Five mice were used for each group and results are the mean of three independent experiments. Student's t-test, *p

    Article Snippet: Reagents Human recombinant NGF and VEGF, neutralizing antibodies against NGF, VEGF and the corresponding isotype control antibodies were purchased from R & D Systems.

    Techniques: Angiogenesis Assay, Mouse Assay, Injection, Multiple Displacement Amplification

    Inhibition of VEGF and NGF by specific neutralizing antibodies significantly suppressed the effects of hDPSC transplantation on the MNCV ( a ) and SNCV ( b ). The neutralizing antibodies against VEGF and NGF were continuously administered using an osmotic pump. The pump was inserted on the day of hDPSC transplantation. The results are expressed as the mean ± SD ( n = 6). ** P

    Journal: Stem Cell Research & Therapy

    Article Title: Transplantation of human dental pulp stem cells ameliorates diabetic polyneuropathy in streptozotocin-induced diabetic nude mice: the role of angiogenic and neurotrophic factors

    doi: 10.1186/s13287-020-01758-9

    Figure Lengend Snippet: Inhibition of VEGF and NGF by specific neutralizing antibodies significantly suppressed the effects of hDPSC transplantation on the MNCV ( a ) and SNCV ( b ). The neutralizing antibodies against VEGF and NGF were continuously administered using an osmotic pump. The pump was inserted on the day of hDPSC transplantation. The results are expressed as the mean ± SD ( n = 6). ** P

    Article Snippet: After 24 h of incubation, the culture media were collected, and the protein concentrations of human VEGF and human NGF in serum and hDPSC culture supernatants were measured by ELISA kits (R & D Systems and Bioscience, Thebarton, South Australia) according to the manufacturer’s instructions.

    Techniques: Inhibition, Transplantation Assay

    Localization of transplanted hDPSCs and expression of angiogenic and neurotrophic factors. a Four weeks after transplantation in the hindlimb skeletal muscles, the transplanted cells were stained with an antibody specific for human nuclei. Bar = 25 μm. b The mRNA expression of VEGF and NGF in the hindlimb skeletal muscles was assessed by real-time quantitative polymerase chain reaction with human probes. The products were observed via agarose gel electrophoresis with ethidium bromide staining

    Journal: Stem Cell Research & Therapy

    Article Title: Transplantation of human dental pulp stem cells ameliorates diabetic polyneuropathy in streptozotocin-induced diabetic nude mice: the role of angiogenic and neurotrophic factors

    doi: 10.1186/s13287-020-01758-9

    Figure Lengend Snippet: Localization of transplanted hDPSCs and expression of angiogenic and neurotrophic factors. a Four weeks after transplantation in the hindlimb skeletal muscles, the transplanted cells were stained with an antibody specific for human nuclei. Bar = 25 μm. b The mRNA expression of VEGF and NGF in the hindlimb skeletal muscles was assessed by real-time quantitative polymerase chain reaction with human probes. The products were observed via agarose gel electrophoresis with ethidium bromide staining

    Article Snippet: After 24 h of incubation, the culture media were collected, and the protein concentrations of human VEGF and human NGF in serum and hDPSC culture supernatants were measured by ELISA kits (R & D Systems and Bioscience, Thebarton, South Australia) according to the manufacturer’s instructions.

    Techniques: Expressing, Transplantation Assay, Staining, Real-time Polymerase Chain Reaction, Agarose Gel Electrophoresis

    VEGF pellet implantation induces blood and lymphatic vessel formation In vivo observation of angiogenesis and lymphangiogenesis in a Prox1-GFP/Flk1::myr-mCherry mouse over 10 days following 150ng VEGF pellet implantation (left 3 columns) or control PBS pellet implantation (far right column). ( A-P ) SteREO Lumar microscopy images of GFP-expressing lymphatic vessels (green), mCherry-expressing blood vessels (red) and overlays (right two columns): ( A-D ) prior to implantation, ( E-H ) 3 days post-implantation, ( I-L ) 7 days post-implantation, and ( MP ) 10 days post-implantation. ( Q-T ) Confocal imaging of the same corneas on day 10. Scale bar: 500 μm in ( A-P ) and 200 μm in ( Q-T ). Arrows ( A ): regularly spaced lymphatic vessels penetrating the cornea in the uninjured eye; arrowheads ( E ): new lymphatic vessels budding from the cornea 3 days after VEGF pellet implantation; asterisk ( I ): one potential new lymphatic vessel budding from the cornea on day 7 after the initial phase; circle ( T ): outline of the implanted PBS control pellet.

    Journal: The FEBS journal

    Article Title: Simultaneous in vivo imaging of blood and lymphatic vessel growth in Prox1-GFP/Flk1::myr-mCherry mice

    doi: 10.1111/febs.13234

    Figure Lengend Snippet: VEGF pellet implantation induces blood and lymphatic vessel formation In vivo observation of angiogenesis and lymphangiogenesis in a Prox1-GFP/Flk1::myr-mCherry mouse over 10 days following 150ng VEGF pellet implantation (left 3 columns) or control PBS pellet implantation (far right column). ( A-P ) SteREO Lumar microscopy images of GFP-expressing lymphatic vessels (green), mCherry-expressing blood vessels (red) and overlays (right two columns): ( A-D ) prior to implantation, ( E-H ) 3 days post-implantation, ( I-L ) 7 days post-implantation, and ( MP ) 10 days post-implantation. ( Q-T ) Confocal imaging of the same corneas on day 10. Scale bar: 500 μm in ( A-P ) and 200 μm in ( Q-T ). Arrows ( A ): regularly spaced lymphatic vessels penetrating the cornea in the uninjured eye; arrowheads ( E ): new lymphatic vessels budding from the cornea 3 days after VEGF pellet implantation; asterisk ( I ): one potential new lymphatic vessel budding from the cornea on day 7 after the initial phase; circle ( T ): outline of the implanted PBS control pellet.

    Article Snippet: Pellets contained 10% (w/v) sulfcralfate (Sigma-Aldrich, St. Louis, MO), 12% (w/v) poly-hydroxyethylmethacrylate (HEMA; Sigma-Aldrich) dissolved in absolute ethanol, and recombinant human VEGF-A (R & D Systems, Minneapolis, MN), bFGF, or PBS.

    Techniques: In Vivo, Microscopy, Expressing, Imaging

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

    Journal: Scientific Reports

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

    doi: 10.1038/s41598-017-09149-6

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

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

    Techniques: TBARS Assay

    pEVs inhibit angiogenesis in vivo . (a) 12-week-old male C57BL/6J mice were subcutaneously injected with either 5 µg of cEVs and pEVs, mixed with Matrigel Matrix supplemented with VEGF 100 ng/mL (450-32 Peprotech) and Heparin 50 units/mL. After 7 days, plugs were harvested. For haemoglobin quantification, plugs were processed by TissueLyser and the haemoglobin content was measured using Drabkin’s reagent kit 525 (Sigma-Aldrich). Each value was first normalized on the total plug protein quantity, measured by BCA assay, and then on the negative control (plugs with vehicle). (b, c) 1-day-old C57BL/6J mouse pups were intraperitoneally injected with a total of 10 μg of cEVs or pEVs, using PBS as control. Mice were sacrificed for retina collection. Dissected retina were stained with isolectin-b4 (green) and digital images were captured using inverted fluorescence confocal microscope. Analyses of the relative radial expansion and of the relative branching point were performed. All data are expressed as means ± SEM, normalized on control (mice treated with vehicle) (n = 8 mice/group). Ordinary one-way ANOVA; * P

    Journal: Journal of Extracellular Vesicles

    Article Title: CD73+ extracellular vesicles inhibit angiogenesis through adenosine A2B receptor signalling

    doi: 10.1080/20013078.2020.1757900

    Figure Lengend Snippet: pEVs inhibit angiogenesis in vivo . (a) 12-week-old male C57BL/6J mice were subcutaneously injected with either 5 µg of cEVs and pEVs, mixed with Matrigel Matrix supplemented with VEGF 100 ng/mL (450-32 Peprotech) and Heparin 50 units/mL. After 7 days, plugs were harvested. For haemoglobin quantification, plugs were processed by TissueLyser and the haemoglobin content was measured using Drabkin’s reagent kit 525 (Sigma-Aldrich). Each value was first normalized on the total plug protein quantity, measured by BCA assay, and then on the negative control (plugs with vehicle). (b, c) 1-day-old C57BL/6J mouse pups were intraperitoneally injected with a total of 10 μg of cEVs or pEVs, using PBS as control. Mice were sacrificed for retina collection. Dissected retina were stained with isolectin-b4 (green) and digital images were captured using inverted fluorescence confocal microscope. Analyses of the relative radial expansion and of the relative branching point were performed. All data are expressed as means ± SEM, normalized on control (mice treated with vehicle) (n = 8 mice/group). Ordinary one-way ANOVA; * P

    Article Snippet: Matrigel plug assay Anesthetized, 12-week-old, male, C57BL/6 N mice were subcutaneously injected in the dorsal back with 5 µg EVs, mixed with 400 µL Matrigel (354234 Corning) supplemented with 100 ng/mL VEGF (450–32 Peprotech) and 50 units/mL Heparin.

    Techniques: In Vivo, Mouse Assay, Injection, BIA-KA, Negative Control, Staining, Fluorescence, Microscopy

    pEVs affect endothelial cell migration and matrix remodelling in vitro . (a, b) Scratch assay was made by scratching a line across the bottom of the dish on a confluent SVEC4-10 monolayer. Cells were treated for 6 h with the medium (Vehicle: DMEM low Glu, 1%P/S, 1% L-Glu; Positive control: Vehicle plus 50 ng/mL VEGF) and 1.75 μg/mL of either cEVs or pEVs. The closure of the wound was quantified with the difference between initial and final scratched areas (Migration Index), calculated by ImageJ and normalized on the vehicle without VEGF. (c, d) For the tube-formation assay SVEC4-10 were seeded in a Matrigel-coated well in the presence of EVs (1.75 μg/mL), using the medium as control (Vehicle: DMEM low Glu, 1%p-s, 1% L-Glu, 10% FBS). After 6 h of incubation, cells were imaged and the quantification of segment length was performed using the ImageJ Angiogenesis Analyze plugin, and normalized on the vehicle. (e, f) The effect on the ECM remodelling was analysed by the FITC-gelatin assay. SVEC4-10 were seeded on the top of the gelatin-coated dishes and cultured for 6hrs with the medium supplemented with 50 ng/mL VEGF in the presence or not of 1.75 μg/mL of either cEVs or pEVs. The vehicle (DMEM low Glu, 1%P/S, 1% L-Glu) was used as control. Cells were then imaged by confocal microscopy and the quantification of the black-digested area (indicated by the with arrows), measured by ImageJ. (g) Analysis of the MMP9/12 activity in the presence of pEVs with InnoZyme MMP2/MMP9 activity assay kit (Millipore) following manufacturer’s instructions. Data are expressed as means ± SEM (3 independent experiments). Kruskal–Wallis test with Dunn’s multiple comparisons test; * P

    Journal: Journal of Extracellular Vesicles

    Article Title: CD73+ extracellular vesicles inhibit angiogenesis through adenosine A2B receptor signalling

    doi: 10.1080/20013078.2020.1757900

    Figure Lengend Snippet: pEVs affect endothelial cell migration and matrix remodelling in vitro . (a, b) Scratch assay was made by scratching a line across the bottom of the dish on a confluent SVEC4-10 monolayer. Cells were treated for 6 h with the medium (Vehicle: DMEM low Glu, 1%P/S, 1% L-Glu; Positive control: Vehicle plus 50 ng/mL VEGF) and 1.75 μg/mL of either cEVs or pEVs. The closure of the wound was quantified with the difference between initial and final scratched areas (Migration Index), calculated by ImageJ and normalized on the vehicle without VEGF. (c, d) For the tube-formation assay SVEC4-10 were seeded in a Matrigel-coated well in the presence of EVs (1.75 μg/mL), using the medium as control (Vehicle: DMEM low Glu, 1%p-s, 1% L-Glu, 10% FBS). After 6 h of incubation, cells were imaged and the quantification of segment length was performed using the ImageJ Angiogenesis Analyze plugin, and normalized on the vehicle. (e, f) The effect on the ECM remodelling was analysed by the FITC-gelatin assay. SVEC4-10 were seeded on the top of the gelatin-coated dishes and cultured for 6hrs with the medium supplemented with 50 ng/mL VEGF in the presence or not of 1.75 μg/mL of either cEVs or pEVs. The vehicle (DMEM low Glu, 1%P/S, 1% L-Glu) was used as control. Cells were then imaged by confocal microscopy and the quantification of the black-digested area (indicated by the with arrows), measured by ImageJ. (g) Analysis of the MMP9/12 activity in the presence of pEVs with InnoZyme MMP2/MMP9 activity assay kit (Millipore) following manufacturer’s instructions. Data are expressed as means ± SEM (3 independent experiments). Kruskal–Wallis test with Dunn’s multiple comparisons test; * P

    Article Snippet: Matrigel plug assay Anesthetized, 12-week-old, male, C57BL/6 N mice were subcutaneously injected in the dorsal back with 5 µg EVs, mixed with 400 µL Matrigel (354234 Corning) supplemented with 100 ng/mL VEGF (450–32 Peprotech) and 50 units/mL Heparin.

    Techniques: Migration, In Vitro, Wound Healing Assay, Positive Control, Tube Formation Assay, Incubation, Cell Culture, Confocal Microscopy, Activity Assay

    One-day post–spinal cord injury (SCI) samples preincubated with vascular endothelial growth factor (VEGF) antibody. ( A ) Western analysis of 24-h samples indicating a reduction in the VEGF165 isoform. ( B ) A Western blot of VEGF165 samples at 2

    Journal:

    Article Title: Reduced Vascular Endothelial Growth Factor Expression in Contusive Spinal Cord Injury

    doi: 10.1089/neu.2008.0779

    Figure Lengend Snippet: One-day post–spinal cord injury (SCI) samples preincubated with vascular endothelial growth factor (VEGF) antibody. ( A ) Western analysis of 24-h samples indicating a reduction in the VEGF165 isoform. ( B ) A Western blot of VEGF165 samples at 2

    Article Snippet: The expression of the VEGF165 isoform was confirmed by performing a preincubation of the 24-h sample with a recombinant human VEGF165 (catalog no. 293-VE; R & D System), with a volume ratio of 1:10, respectively.

    Techniques: Western Blot

    Temporal vascular endothelial growth factor (VEGF) expression profile in regions around the lesion epicenter after spinal cord injury (SCI) compared to sham controls. VEGF165 isoform was significantly reduced 1 day after injury and remained reduced out

    Journal:

    Article Title: Reduced Vascular Endothelial Growth Factor Expression in Contusive Spinal Cord Injury

    doi: 10.1089/neu.2008.0779

    Figure Lengend Snippet: Temporal vascular endothelial growth factor (VEGF) expression profile in regions around the lesion epicenter after spinal cord injury (SCI) compared to sham controls. VEGF165 isoform was significantly reduced 1 day after injury and remained reduced out

    Article Snippet: The expression of the VEGF165 isoform was confirmed by performing a preincubation of the 24-h sample with a recombinant human VEGF165 (catalog no. 293-VE; R & D System), with a volume ratio of 1:10, respectively.

    Techniques: Expressing

    Effect of miR-126-5p on HUVEC proliferation and downstream signaling pathways. ( A ) qRT-PCR results showing the expression of miR-126-5p in the different groups. ( B ) Representative western blot showing the expression of p-Akt, VEGF, eNOS and CD31 in each group (normalized to the expression of β-tubulin). ( C ) Densitometry analyses of p-Akt, VEGF, eNOS and CD31 expression in each group normalized to the expression of t-Akt and β-tubulin. The error bars represent the ±SDs. *P

    Journal: Aging (Albany NY)

    Article Title: Increasing the expression of microRNA-126-5p in the temporal muscle can promote angiogenesis in the chronically ischemic brains of rats subjected to two-vessel occlusion plus encephalo-myo-synangiosis

    doi: 10.18632/aging.103431

    Figure Lengend Snippet: Effect of miR-126-5p on HUVEC proliferation and downstream signaling pathways. ( A ) qRT-PCR results showing the expression of miR-126-5p in the different groups. ( B ) Representative western blot showing the expression of p-Akt, VEGF, eNOS and CD31 in each group (normalized to the expression of β-tubulin). ( C ) Densitometry analyses of p-Akt, VEGF, eNOS and CD31 expression in each group normalized to the expression of t-Akt and β-tubulin. The error bars represent the ±SDs. *P

    Article Snippet: Subsequently, the fixed sections were washed and incubated for 1 h with primary antibodies against vWF (1:200; Cell Signaling Technology, Shanghai, China), VEGF (1:200; Cell Signaling Technology), CD31 (1:100; Santa Cruz Biotechnology, Dallas, TX, USA), or eNOS (1:200; Cell Signaling Technology) overnight at 4°C.

    Techniques: Quantitative RT-PCR, Expressing, Western Blot

    Effectiveness of the 2VO+EMS model in promoting EC proliferation determined by immunofluorescence and western blot assays. ( A ) Brain samples from 2VO+EMS rats (the black arrow indicates the adhered TM tissue). ( B , C ) Hematoxylin and eosin (HE) and immunofluorescence results showing that there was a significantly higher number of vWF(+) cells in the TM tissue on the EMS side than on the non-EMS side. Bar = 20 μm. ( D ) HE-stained slide of a brain from a 2VO+EMS rat. The two black frames indicate the portion of the brain tissue in contact with the TM and the brain tissue on the symmetrical part of the 2VO side. Bar = 1 mm. Enlarged image of the black frame on ( E ) the non-EMS side and ( F ) the EMS side. Bar = 200 μm. VEGF(+) immunofluorescence results for ( G ) the non-EMS side and ( H ) the EMS side. Bar = 200 μm. The yellow curve indicates the brain surface involved in EMS. Immunofluorescence ( I , J ) and western blot ( K , L ) results showing that there was significantly higher VEGF expression on the EMS side in the 2VO+EMS rat brains than on the non-EMS side. Bar = 20 μm. The error bars represent the ±SDs. VEGF: vascular endothelial growth factor; EMS: encephalo-myo-synangiosis; EC: endothelial cell; vWF: von Willebrand factor.

    Journal: Aging (Albany NY)

    Article Title: Increasing the expression of microRNA-126-5p in the temporal muscle can promote angiogenesis in the chronically ischemic brains of rats subjected to two-vessel occlusion plus encephalo-myo-synangiosis

    doi: 10.18632/aging.103431

    Figure Lengend Snippet: Effectiveness of the 2VO+EMS model in promoting EC proliferation determined by immunofluorescence and western blot assays. ( A ) Brain samples from 2VO+EMS rats (the black arrow indicates the adhered TM tissue). ( B , C ) Hematoxylin and eosin (HE) and immunofluorescence results showing that there was a significantly higher number of vWF(+) cells in the TM tissue on the EMS side than on the non-EMS side. Bar = 20 μm. ( D ) HE-stained slide of a brain from a 2VO+EMS rat. The two black frames indicate the portion of the brain tissue in contact with the TM and the brain tissue on the symmetrical part of the 2VO side. Bar = 1 mm. Enlarged image of the black frame on ( E ) the non-EMS side and ( F ) the EMS side. Bar = 200 μm. VEGF(+) immunofluorescence results for ( G ) the non-EMS side and ( H ) the EMS side. Bar = 200 μm. The yellow curve indicates the brain surface involved in EMS. Immunofluorescence ( I , J ) and western blot ( K , L ) results showing that there was significantly higher VEGF expression on the EMS side in the 2VO+EMS rat brains than on the non-EMS side. Bar = 20 μm. The error bars represent the ±SDs. VEGF: vascular endothelial growth factor; EMS: encephalo-myo-synangiosis; EC: endothelial cell; vWF: von Willebrand factor.

    Article Snippet: Subsequently, the fixed sections were washed and incubated for 1 h with primary antibodies against vWF (1:200; Cell Signaling Technology, Shanghai, China), VEGF (1:200; Cell Signaling Technology), CD31 (1:100; Santa Cruz Biotechnology, Dallas, TX, USA), or eNOS (1:200; Cell Signaling Technology) overnight at 4°C.

    Techniques: Immunofluorescence, Western Blot, Staining, Expressing

    Western blot results showing the expression of relevant proteins in each group. ( A ) Representative western blot showing the expression of VEGF, CD31, eNOS and p-Akt in the ischemic brain tissue adjacent to the TM in each group (normalized to β-tubulin expression). ( B ) Densitometry analyses of VEGF, CD31, eNOS and p-Akt expression normalized to the expression of β-tubulin and t-Akt. The error bars represent the ±SDs. *P

    Journal: Aging (Albany NY)

    Article Title: Increasing the expression of microRNA-126-5p in the temporal muscle can promote angiogenesis in the chronically ischemic brains of rats subjected to two-vessel occlusion plus encephalo-myo-synangiosis

    doi: 10.18632/aging.103431

    Figure Lengend Snippet: Western blot results showing the expression of relevant proteins in each group. ( A ) Representative western blot showing the expression of VEGF, CD31, eNOS and p-Akt in the ischemic brain tissue adjacent to the TM in each group (normalized to β-tubulin expression). ( B ) Densitometry analyses of VEGF, CD31, eNOS and p-Akt expression normalized to the expression of β-tubulin and t-Akt. The error bars represent the ±SDs. *P

    Article Snippet: Subsequently, the fixed sections were washed and incubated for 1 h with primary antibodies against vWF (1:200; Cell Signaling Technology, Shanghai, China), VEGF (1:200; Cell Signaling Technology), CD31 (1:100; Santa Cruz Biotechnology, Dallas, TX, USA), or eNOS (1:200; Cell Signaling Technology) overnight at 4°C.

    Techniques: Western Blot, Expressing

    Expression of CD31 and VEGF in TM tissues from each group. ( A ) HE and immunofluorescence results showing CD31 and VEGF expression in TM tissues from each group. Bar = 20 μm. ( B ) Quantification of CD31(+), VEGF(+), and CD31/VEGF(+) cells in TM tissue. The data are reported as the means ± SDs. n = 8. VEGF: vascular endothelial growth factor.

    Journal: Aging (Albany NY)

    Article Title: Increasing the expression of microRNA-126-5p in the temporal muscle can promote angiogenesis in the chronically ischemic brains of rats subjected to two-vessel occlusion plus encephalo-myo-synangiosis

    doi: 10.18632/aging.103431

    Figure Lengend Snippet: Expression of CD31 and VEGF in TM tissues from each group. ( A ) HE and immunofluorescence results showing CD31 and VEGF expression in TM tissues from each group. Bar = 20 μm. ( B ) Quantification of CD31(+), VEGF(+), and CD31/VEGF(+) cells in TM tissue. The data are reported as the means ± SDs. n = 8. VEGF: vascular endothelial growth factor.

    Article Snippet: Subsequently, the fixed sections were washed and incubated for 1 h with primary antibodies against vWF (1:200; Cell Signaling Technology, Shanghai, China), VEGF (1:200; Cell Signaling Technology), CD31 (1:100; Santa Cruz Biotechnology, Dallas, TX, USA), or eNOS (1:200; Cell Signaling Technology) overnight at 4°C.

    Techniques: Expressing, Immunofluorescence

    Endogenous neurogenesis was increased in the NSC-transplanted Tg2576 mice. ( a ) At 2.5 months after transplantation, endogenous neurogenesis was observed in the DG of hippocampus by immunohistochemisty using anti-DCX antibody. DCX-positive cells were calculated in mm 2 area of DG. Graph represents that endogenous neurogenesis was enhanced by NSC transplantation. ( b ) The levels of PSA-NCAM were analyzed by immunohistochemistry. In Tg-NSC group, the levels of PSA-NCAM were increased compared with the Tg-sham group. Graph represents that PSA-NCAM was enhanced by NSC transplantation. ( c ) In Tg-NSC group, the levels of PSA-NCAM and VEGF were increased compared with the Tg-sham group. Quantitative analysis shows that NSC transplantation significantly increased the level of PSA-NCAM and VEGF expressions. * P

    Journal: Cell Death & Disease

    Article Title: Neural stem cell transplantation at critical period improves learning and memory through restoring synaptic impairment in Alzheimer's disease mouse model

    doi: 10.1038/cddis.2015.138

    Figure Lengend Snippet: Endogenous neurogenesis was increased in the NSC-transplanted Tg2576 mice. ( a ) At 2.5 months after transplantation, endogenous neurogenesis was observed in the DG of hippocampus by immunohistochemisty using anti-DCX antibody. DCX-positive cells were calculated in mm 2 area of DG. Graph represents that endogenous neurogenesis was enhanced by NSC transplantation. ( b ) The levels of PSA-NCAM were analyzed by immunohistochemistry. In Tg-NSC group, the levels of PSA-NCAM were increased compared with the Tg-sham group. Graph represents that PSA-NCAM was enhanced by NSC transplantation. ( c ) In Tg-NSC group, the levels of PSA-NCAM and VEGF were increased compared with the Tg-sham group. Quantitative analysis shows that NSC transplantation significantly increased the level of PSA-NCAM and VEGF expressions. * P

    Article Snippet: Antibodies Primary antibodies were used as follows: goat anti-DCX (1 : 100; Santa Cruz, Dallas, TX, USA), mouse anti-MAP2 (1 : 100; Millipore, Billerica, MA, USA), rabbit anti-Iba1 (1 : 2000; Wako, Richmond, VA, USA), mouse anti-6E10 (1 : 1000, Covance, San Diego, CA, USA), mouse anti-phospho tau (1 : 1000, Pierce, Rockford, IL, USA), goat anti-tau (C17) (1 : 1000, Santa Cruz), rat anti-neprilysin (1 : 500, R & D Systems, Inc.), mouse anti PSA-NCAM (1 : 2000, Millipore), rabbit anti-VEGF (1 : 1000, Santa Cruz), mouse anti-PSD-95 (1 : 2000, Thermo Scientific, Waltham, MA, USA), rabbit anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH; 1 : 10000, Ab Frontier, Seoul, South Korea).

    Techniques: Mouse Assay, Transplantation Assay, Immunohistochemistry

    SLPI promotes proliferation of adult neural stem cells and induces cyclin D1 . A: Proliferation of rat neural stem cells after treatment with SLPI or VEGF, respectively. Cells were treated with indicated amounts of SLPI or VEGF for three days. Afterwards, they were pulsed with 10 μM BrdU. Proportion of BrdU-positive cells (+ std dev) was determined with the FITC BrdU Flow Kit. Presentation of a representative result of three experiments. *:p

    Journal: Journal of Neuroinflammation

    Article Title: Novel role for SLPI in MOG-induced EAE revealed by spinal cord expression analysis

    doi: 10.1186/1742-2094-5-20

    Figure Lengend Snippet: SLPI promotes proliferation of adult neural stem cells and induces cyclin D1 . A: Proliferation of rat neural stem cells after treatment with SLPI or VEGF, respectively. Cells were treated with indicated amounts of SLPI or VEGF for three days. Afterwards, they were pulsed with 10 μM BrdU. Proportion of BrdU-positive cells (+ std dev) was determined with the FITC BrdU Flow Kit. Presentation of a representative result of three experiments. *:p

    Article Snippet: After one day the specified amounts of recombinant SLPI, recombinant VEGF (R & D Systems) or recombinant α1 -antitrypsin (α1 -AT, Sigma Aldrich) were added.

    Techniques: Flow Cytometry

    Long-term effects of intravitreally injected anti-VEGF antibody on BAT. (A) Quantitative analyses of the number of large lipid droplets ( > 50 μm 2 ) per field at x400 magnification ( n = 3–6). The effects of anti-VEGF antibody were quantitatively analyzed by comparison to the group treated with intravitreal PBS injection as 100%. (B) Quantitative analyses of vascularity of interscapular BAT demonstrated by isolectin B4 staining ( n = 3–6). The effects of anti-VEGF antibody were quantitatively analyzed by comparison to the group treated with intravitreal PBS injection as 100%. (C) The changes in body weight from P14 to P56. Anti-VEGF, anti-VEGF antibody. NS , not significant (two-tailed, unpaired T-test).

    Journal: PLoS ONE

    Article Title: Intravitreally Injected Anti-VEGF Antibody Reduces Brown Fat in Neonatal Mice

    doi: 10.1371/journal.pone.0134308

    Figure Lengend Snippet: Long-term effects of intravitreally injected anti-VEGF antibody on BAT. (A) Quantitative analyses of the number of large lipid droplets ( > 50 μm 2 ) per field at x400 magnification ( n = 3–6). The effects of anti-VEGF antibody were quantitatively analyzed by comparison to the group treated with intravitreal PBS injection as 100%. (B) Quantitative analyses of vascularity of interscapular BAT demonstrated by isolectin B4 staining ( n = 3–6). The effects of anti-VEGF antibody were quantitatively analyzed by comparison to the group treated with intravitreal PBS injection as 100%. (C) The changes in body weight from P14 to P56. Anti-VEGF, anti-VEGF antibody. NS , not significant (two-tailed, unpaired T-test).

    Article Snippet: To estimate the level of anti-VEGF antibody using goat IgG ELISA, we utilized PBS as the control.

    Techniques: Injection, Staining, Two Tailed Test

    Ocular and systemic consequences of intravitreally injected anti-VEGF antibody. (A) Effects of intravitreally injected anti-VEGF antibody (1 μg/eye) on retinal neovascularization in OIR mice ( n = 6). Neovascular tufts were highlighted with yellow pseudocolor on representative images of isolectin B4-stained retina. The area of neovascular tufts was normalized to total retinal area; then, the effects of anti-VEGF antibody were quantified and normalized to the control (intravitreal PBS injection). Scale bar, 200 μm. (B) Retinal VEGF concentrations at P17 with intravitreal injection of PBS or anti-VEGF antibody ( n = 3). The level of VEGF was normalized to total amounts of proteins in the retina. (C) Serum concentrations of anti-VEGF antibody after intravitreal injection at P14, P17, and P21 ( n = 3–6). (D) Serum VEGF concentrations after intravitreal injection of anti-VEGF antibody at P17, P21, and P28 ( n = 3–6). Data are presented as mean ± SEM in graphs. Anti-VEGF, anti-VEGF antibody. NS , not significant; **, P

    Journal: PLoS ONE

    Article Title: Intravitreally Injected Anti-VEGF Antibody Reduces Brown Fat in Neonatal Mice

    doi: 10.1371/journal.pone.0134308

    Figure Lengend Snippet: Ocular and systemic consequences of intravitreally injected anti-VEGF antibody. (A) Effects of intravitreally injected anti-VEGF antibody (1 μg/eye) on retinal neovascularization in OIR mice ( n = 6). Neovascular tufts were highlighted with yellow pseudocolor on representative images of isolectin B4-stained retina. The area of neovascular tufts was normalized to total retinal area; then, the effects of anti-VEGF antibody were quantified and normalized to the control (intravitreal PBS injection). Scale bar, 200 μm. (B) Retinal VEGF concentrations at P17 with intravitreal injection of PBS or anti-VEGF antibody ( n = 3). The level of VEGF was normalized to total amounts of proteins in the retina. (C) Serum concentrations of anti-VEGF antibody after intravitreal injection at P14, P17, and P21 ( n = 3–6). (D) Serum VEGF concentrations after intravitreal injection of anti-VEGF antibody at P17, P21, and P28 ( n = 3–6). Data are presented as mean ± SEM in graphs. Anti-VEGF, anti-VEGF antibody. NS , not significant; **, P

    Article Snippet: To estimate the level of anti-VEGF antibody using goat IgG ELISA, we utilized PBS as the control.

    Techniques: Injection, Mouse Assay, Staining

    Effects of intravitreally injected anti-VEGF antibody on BAT of neonatal mice. (A) Concentrations of VEGF in interscapular BAT at P21 and P28. The level of VEGF was normalized to total amounts of proteins in BAT ( n = 3–6). (B) Representative images of H E staining of interscapular BAT after intravitreal injection of PBS or anti-VEGF antibody show enlarged lipid droplets. Scale bar, 20 μm. (C) Quantitative analyses of vascularity of interscapular BAT based on isolectin B4 staining ( n = 3–6). The effects of anti-VEGF antibody were quantified and normalized to the control (intravitreal PBS injection). (D) Relative expression of Ucp1 and Ppargc1a in interscapular BAT ( n = 3–6). Data are presented as mean ± SEM in graphs. Anti-VEGF, anti-VEGF antibody. *, P

    Journal: PLoS ONE

    Article Title: Intravitreally Injected Anti-VEGF Antibody Reduces Brown Fat in Neonatal Mice

    doi: 10.1371/journal.pone.0134308

    Figure Lengend Snippet: Effects of intravitreally injected anti-VEGF antibody on BAT of neonatal mice. (A) Concentrations of VEGF in interscapular BAT at P21 and P28. The level of VEGF was normalized to total amounts of proteins in BAT ( n = 3–6). (B) Representative images of H E staining of interscapular BAT after intravitreal injection of PBS or anti-VEGF antibody show enlarged lipid droplets. Scale bar, 20 μm. (C) Quantitative analyses of vascularity of interscapular BAT based on isolectin B4 staining ( n = 3–6). The effects of anti-VEGF antibody were quantified and normalized to the control (intravitreal PBS injection). (D) Relative expression of Ucp1 and Ppargc1a in interscapular BAT ( n = 3–6). Data are presented as mean ± SEM in graphs. Anti-VEGF, anti-VEGF antibody. *, P

    Article Snippet: To estimate the level of anti-VEGF antibody using goat IgG ELISA, we utilized PBS as the control.

    Techniques: Injection, Mouse Assay, Staining, Expressing

    Neurotrophic factor expression in mesenchymal stem cells. ( A ) RT-PCR analysis of neurotrophic factor transcripts NGF, GDNF, NT-3, BDNF, VEGF from undifferentiated MSC (uMSC) and differentiated MSC (dMSC) harvested from young and old donors. Amplicon size is shown in base pairs (bp). ( B and C ) ELISA was used to determine BDNF and VEGF protein levels in cell culture supernatants. ***P

    Journal: PLoS ONE

    Article Title: Aging Effect on Neurotrophic Activity of Human Mesenchymal Stem Cells

    doi: 10.1371/journal.pone.0045052

    Figure Lengend Snippet: Neurotrophic factor expression in mesenchymal stem cells. ( A ) RT-PCR analysis of neurotrophic factor transcripts NGF, GDNF, NT-3, BDNF, VEGF from undifferentiated MSC (uMSC) and differentiated MSC (dMSC) harvested from young and old donors. Amplicon size is shown in base pairs (bp). ( B and C ) ELISA was used to determine BDNF and VEGF protein levels in cell culture supernatants. ***P

    Article Snippet: Other inserts contained either 10 µg/ml anti-BDNF antibody (Millipore) or 1 µg/ml anti-VEGF antibody (R & D Systems, UK).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Amplification, Enzyme-linked Immunosorbent Assay, Cell Culture