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    Name:
    Recombinant Murine EG VEGF
    Description:
    EG VEGF is a secreted angiogenetic mitogen growth factor expressed in the steroidogenic glands ovary testis adrenal and placenta EG VEGF induces proliferation and migration fenestration formation of membrane discontinuities in capillary endothelial cells derived from endocrine glands The murine EG VEGF gene codes for a 105 amino acid polypeptide containing an N terminal signal sequence of 19 amino acids Recombinant murine EG VEGF is a 9 6 kDa protein consisting of 86 amino acid residues and ten cysteine residues that potentially form five pairs of intra molecular disulfide bonds
    Catalog Number:
    315-29-100UG
    Price:
    600.00
    Category:
    Recombinant Proteins
    Source:
    E.coli
    Purity:
    98.0
    Quantity:
    100UG
    Buy from Supplier


    Structured Review

    PeproTech vegf
    Quantification of asthma-related mediators in the BALF. The levels of pro-inflammatory T H 2 cytokines, including ( A ) IL-4 and ( B ) IL-13, ( C ) an anti-inflammatory cytokine, IL-10, and profibrotic cytokines, including ( D ) <t>VEGF</t> and ( E ) <t>TGF-β,</t> were quantified by ELISA ( n = 6 mice per group).
    EG VEGF is a secreted angiogenetic mitogen growth factor expressed in the steroidogenic glands ovary testis adrenal and placenta EG VEGF induces proliferation and migration fenestration formation of membrane discontinuities in capillary endothelial cells derived from endocrine glands The murine EG VEGF gene codes for a 105 amino acid polypeptide containing an N terminal signal sequence of 19 amino acids Recombinant murine EG VEGF is a 9 6 kDa protein consisting of 86 amino acid residues and ten cysteine residues that potentially form five pairs of intra molecular disulfide bonds
    https://www.bioz.com/result/vegf/product/PeproTech
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    vegf - by Bioz Stars, 2021-04
    95/100 stars

    Images

    1) Product Images from "Nanoparticle-based thymulin gene therapy therapeutically reverses key pathology of experimental allergic asthma"

    Article Title: Nanoparticle-based thymulin gene therapy therapeutically reverses key pathology of experimental allergic asthma

    Journal: Science Advances

    doi: 10.1126/sciadv.aay7973

    Quantification of asthma-related mediators in the BALF. The levels of pro-inflammatory T H 2 cytokines, including ( A ) IL-4 and ( B ) IL-13, ( C ) an anti-inflammatory cytokine, IL-10, and profibrotic cytokines, including ( D ) VEGF and ( E ) TGF-β, were quantified by ELISA ( n = 6 mice per group).
    Figure Legend Snippet: Quantification of asthma-related mediators in the BALF. The levels of pro-inflammatory T H 2 cytokines, including ( A ) IL-4 and ( B ) IL-13, ( C ) an anti-inflammatory cytokine, IL-10, and profibrotic cytokines, including ( D ) VEGF and ( E ) TGF-β, were quantified by ELISA ( n = 6 mice per group).

    Techniques Used: Enzyme-linked Immunosorbent Assay, Mouse Assay

    2) Product Images from "Dynamic interaction networks in a hierarchically organized tissue"

    Article Title: Dynamic interaction networks in a hierarchically organized tissue

    Journal: Molecular Systems Biology

    doi: 10.1038/msb.2010.71

    Integration of endogenous regulatory signals in the HSC intracellular self-renewal network. ( A ) Endogenous secreted stimulators (VEGF, EGF, PDGF, and 5HT1) and inhibitors (CCL3, CCL4, CXCL10, TNFSF9, and TGFB2) activate cell surface receptors on HSCs, inducing signal transduction events, which are coherently processed by the intracellular network to modulate rates of self-renewal versus differentiation. Common signal transduction molecules shared by stimulatory pathways (left; green box), inhibitory pathways (right; red box), and both (center) are densely connected to known self-renewal effector genes. Physical protein–protein interactions from stimulatory and inhibitory pathways are represented as green and red edges, respectively, whereas internal interactions are represented as blue edges. ( B ) Five small molecule antagonists, described in the table with targets indicated by numbers on the network, were tested for functional effects on 8-day fold expansions of total cells (TNC), progenitors (CFC), and primitive progenitors (LTCIC) with respect to control cultures. ( C ) To classify the functional activities of the molecules, culture simulations were run over a feasible range of HSC self-renewal probabilities and proliferation rates. On the basis of ΔWRSS ranking of effects on TNC, CFC, and LTCIC output, inhibition of PI3K and Raf, reduces self-renewal, whereas inhibition of Akt reduces proliferation. Experimental data is overlaid for visual depiction of the model-based functional classifications of the kinase inhibitors. Boxes indicate ±1 s.d., overlaid at estimated effects levels.
    Figure Legend Snippet: Integration of endogenous regulatory signals in the HSC intracellular self-renewal network. ( A ) Endogenous secreted stimulators (VEGF, EGF, PDGF, and 5HT1) and inhibitors (CCL3, CCL4, CXCL10, TNFSF9, and TGFB2) activate cell surface receptors on HSCs, inducing signal transduction events, which are coherently processed by the intracellular network to modulate rates of self-renewal versus differentiation. Common signal transduction molecules shared by stimulatory pathways (left; green box), inhibitory pathways (right; red box), and both (center) are densely connected to known self-renewal effector genes. Physical protein–protein interactions from stimulatory and inhibitory pathways are represented as green and red edges, respectively, whereas internal interactions are represented as blue edges. ( B ) Five small molecule antagonists, described in the table with targets indicated by numbers on the network, were tested for functional effects on 8-day fold expansions of total cells (TNC), progenitors (CFC), and primitive progenitors (LTCIC) with respect to control cultures. ( C ) To classify the functional activities of the molecules, culture simulations were run over a feasible range of HSC self-renewal probabilities and proliferation rates. On the basis of ΔWRSS ranking of effects on TNC, CFC, and LTCIC output, inhibition of PI3K and Raf, reduces self-renewal, whereas inhibition of Akt reduces proliferation. Experimental data is overlaid for visual depiction of the model-based functional classifications of the kinase inhibitors. Boxes indicate ±1 s.d., overlaid at estimated effects levels.

    Techniques Used: Transduction, Functional Assay, Inhibition

    Schematic summary of experimental findings. In addition to exogenous growth factors (FLT3LG, KITL, and THPO in our cultures), stem cell output is regulated by secreted factor-mediated cell–cell interactions. An antagonistic axis of intercellular communication is established, wherein culture-derived monocytes secrete high levels of factors (CCL3, CCL4, CXCL10, TGFB2, and TNSFS9), which inhibit stem cell self-renewal, whereas culture-derived megakaryocytes secrete high levels of factors (EGF, PDGFB, VEGF, and serotonin (5HT1)), which stimulate stem cell self-renewal, functioning as a coupled positive and negative intercellular feedback circuit.
    Figure Legend Snippet: Schematic summary of experimental findings. In addition to exogenous growth factors (FLT3LG, KITL, and THPO in our cultures), stem cell output is regulated by secreted factor-mediated cell–cell interactions. An antagonistic axis of intercellular communication is established, wherein culture-derived monocytes secrete high levels of factors (CCL3, CCL4, CXCL10, TGFB2, and TNSFS9), which inhibit stem cell self-renewal, whereas culture-derived megakaryocytes secrete high levels of factors (EGF, PDGFB, VEGF, and serotonin (5HT1)), which stimulate stem cell self-renewal, functioning as a coupled positive and negative intercellular feedback circuit.

    Techniques Used: Derivative Assay

    3) Product Images from "Recombinant RGD-disintegrin DisBa-01 blocks integrin αvβ3 and impairs VEGF signaling in endothelial cells"

    Article Title: Recombinant RGD-disintegrin DisBa-01 blocks integrin αvβ3 and impairs VEGF signaling in endothelial cells

    Journal: Cell Communication and Signaling : CCS

    doi: 10.1186/s12964-019-0339-1

    Dis Ba -01 inhibits ERK1/2 and PI3K phosphorylation. HUVECs (5 × 10 5 cells/well) were seeded in 6-well plates and left to adhere at 37 °C, 5% CO 2 , overnight, followed by a period of 24 h of starvation at serum-free medium. Cells were treated with 1 ml of DMEM supplemented with 10% FBS and either Dis Ba -01 (1000 nM), VEGF (10 ng/mL) or a co-treatment and incubated for 1 and 24 h at 37 °C, 5% CO 2 , followed by cell lysis. Twenty micrograms of protein from the cell lysate were separated on SDS-PAGE. Blots were probed with antibodies to a P-TY187 ERK1 + ERK2 and anti-ERK1 + ERK2; to b P-TY607 PI3K and anti-PI3K and GAPDH, this last used to normalize loading. Bands corresponding to all proteins were quantified by densitometry using the ImageJ FIJI program. Bar graph shows the mean ± SE of phosphorylated ERK1 + ERK2/ERK1 + ERK2/GAPDH and PI3K/PI3K/GAPDH expression from three independent experiments. Values of *p
    Figure Legend Snippet: Dis Ba -01 inhibits ERK1/2 and PI3K phosphorylation. HUVECs (5 × 10 5 cells/well) were seeded in 6-well plates and left to adhere at 37 °C, 5% CO 2 , overnight, followed by a period of 24 h of starvation at serum-free medium. Cells were treated with 1 ml of DMEM supplemented with 10% FBS and either Dis Ba -01 (1000 nM), VEGF (10 ng/mL) or a co-treatment and incubated for 1 and 24 h at 37 °C, 5% CO 2 , followed by cell lysis. Twenty micrograms of protein from the cell lysate were separated on SDS-PAGE. Blots were probed with antibodies to a P-TY187 ERK1 + ERK2 and anti-ERK1 + ERK2; to b P-TY607 PI3K and anti-PI3K and GAPDH, this last used to normalize loading. Bands corresponding to all proteins were quantified by densitometry using the ImageJ FIJI program. Bar graph shows the mean ± SE of phosphorylated ERK1 + ERK2/ERK1 + ERK2/GAPDH and PI3K/PI3K/GAPDH expression from three independent experiments. Values of *p

    Techniques Used: Incubation, Lysis, SDS Page, Expressing

    Dis Ba -01 promotes FAK, Src and paxillin phosphorylation. HUVECs (5 × 10 5 cells/well) were seeded in 6-well plates and left to adhere at 37 °C, 5% CO 2 , overnight, followed by a period of 24 h of starvation at serum-free medium. Cells were treated with 1 ml of DMEM supplemented with 10% FBS and either Dis Ba -01 (1000 nM), VEGF (10 ng/mL) or a co-treatment and incubated for 1 and 24 h at 37 °C, 5% CO 2 , followed by cell lysis. Twenty micrograms of protein from the cell lysate were separated on SDS-PAGE. Blots were probed with antibodies to a P-Y397 FAK and anti-FAK; to b P-TY418 Src; to c phospho LIM1 Paxilin; and GAPDH, this last used to normalize loading. Bands corresponding to all proteins were quantified by densitometry using the ImageJ FIJI program. Bar graph shows the mean ± SE of phosphorylated pFAK/FAK/GAPDH, pSrc/GAPDH and pLIM1Paxilin/GAPDH expression from three independent experiments. Values of *p
    Figure Legend Snippet: Dis Ba -01 promotes FAK, Src and paxillin phosphorylation. HUVECs (5 × 10 5 cells/well) were seeded in 6-well plates and left to adhere at 37 °C, 5% CO 2 , overnight, followed by a period of 24 h of starvation at serum-free medium. Cells were treated with 1 ml of DMEM supplemented with 10% FBS and either Dis Ba -01 (1000 nM), VEGF (10 ng/mL) or a co-treatment and incubated for 1 and 24 h at 37 °C, 5% CO 2 , followed by cell lysis. Twenty micrograms of protein from the cell lysate were separated on SDS-PAGE. Blots were probed with antibodies to a P-Y397 FAK and anti-FAK; to b P-TY418 Src; to c phospho LIM1 Paxilin; and GAPDH, this last used to normalize loading. Bands corresponding to all proteins were quantified by densitometry using the ImageJ FIJI program. Bar graph shows the mean ± SE of phosphorylated pFAK/FAK/GAPDH, pSrc/GAPDH and pLIM1Paxilin/GAPDH expression from three independent experiments. Values of *p

    Techniques Used: Incubation, Lysis, SDS Page, Expressing

    DisBa-01 induces morphological changes in endothelial cells. HUVECs (3 × 10 4 cells/well) were plated in a 96-well microplate previously coated with FN (1 μg/cm 2 ), in serum-free DMEM and incubated overnight at 37 °C, 5% CO 2 . Cells were exposed to VEGF (10 ng/mL), Dis Ba -01 (1000 nM) and VEGF plus Dis Ba -01 for 30 min in DMEM 10% FBS. Cell nuclei were stained with DAPI (0.7 ng/μl) and cytoplasm was stained with Alexa Fluor™ 488 phalloidin for 10 min. Images were observed with 60x magnification. Representative images were obtained from three independent experiments. Scale bar = 50 μm (left panel) and 20 μm (right panel)
    Figure Legend Snippet: DisBa-01 induces morphological changes in endothelial cells. HUVECs (3 × 10 4 cells/well) were plated in a 96-well microplate previously coated with FN (1 μg/cm 2 ), in serum-free DMEM and incubated overnight at 37 °C, 5% CO 2 . Cells were exposed to VEGF (10 ng/mL), Dis Ba -01 (1000 nM) and VEGF plus Dis Ba -01 for 30 min in DMEM 10% FBS. Cell nuclei were stained with DAPI (0.7 ng/μl) and cytoplasm was stained with Alexa Fluor™ 488 phalloidin for 10 min. Images were observed with 60x magnification. Representative images were obtained from three independent experiments. Scale bar = 50 μm (left panel) and 20 μm (right panel)

    Techniques Used: Incubation, Staining

    Dis Ba -01 effects on VEGF-induced HUVEC viability, invasion, migration and adhesion. a Cells were treated with Dis Ba -01 (1000 nM), VEGF (10 ng/mL) or both proteins in DMEM supplemented with 0.5% FBS followed by 24 h of incubation. Cell viability was measured by spectrophotometry at 540 nm after incubation with MTT. b HUVECs (2 × 10 5 cells/well) were treated with 1000 nM Dis Ba -01 and/or VEGF (10 ng/mL) on serum-free DMEM for 30 min at 4 °C. Cells were pipetted into the Boyden’s chamber and then it was inserted on well containing DMEM 10% FBS. The negative control comprised of serum-free DMEM on the wells. Invasion was allowed to occur for 18 h at 37 °C. Cell nuclei were stained with DAPI (0.7 ng/μl). Quantification of invasive cells was measured by automated cell counting. c-d For the migration assay, HUVECs (1 × 10 5 cells/well) were exposed to Dis Ba -01 (1, 10, 100 and 1000 nM), VEGF (10 ng/mL) or VEGF plus Dis Ba -01 (1000 nM) and immediately inserted into the Boyden’s chamber. The chambers were immersed in 10% FBS medium and allowed to migrate for 6 h at 37 °C. Control chambers were inserted in serum-free medium. Cell nuclei were stained with DAPI (0.7 ng/μl) and cell migration was measured by automated cell counting. e-f HUVECs (1 × 10 5 cells/well) were treated with Dis Ba -01 (1000 nM) and/or VEGF (10 ng/mL) and were immediately incubated (37 °C, 1 h) in fibronectin and vitronectin precoated-wells. Negative control was comprised of wells coated with 2% BSA. Cell nuclei were stained with DAPI (0.7 ng/μl) and quantification of adhesion cells was measured by automated cell counting. Results represent the average of three independent experiments in triplicate. Values of * p
    Figure Legend Snippet: Dis Ba -01 effects on VEGF-induced HUVEC viability, invasion, migration and adhesion. a Cells were treated with Dis Ba -01 (1000 nM), VEGF (10 ng/mL) or both proteins in DMEM supplemented with 0.5% FBS followed by 24 h of incubation. Cell viability was measured by spectrophotometry at 540 nm after incubation with MTT. b HUVECs (2 × 10 5 cells/well) were treated with 1000 nM Dis Ba -01 and/or VEGF (10 ng/mL) on serum-free DMEM for 30 min at 4 °C. Cells were pipetted into the Boyden’s chamber and then it was inserted on well containing DMEM 10% FBS. The negative control comprised of serum-free DMEM on the wells. Invasion was allowed to occur for 18 h at 37 °C. Cell nuclei were stained with DAPI (0.7 ng/μl). Quantification of invasive cells was measured by automated cell counting. c-d For the migration assay, HUVECs (1 × 10 5 cells/well) were exposed to Dis Ba -01 (1, 10, 100 and 1000 nM), VEGF (10 ng/mL) or VEGF plus Dis Ba -01 (1000 nM) and immediately inserted into the Boyden’s chamber. The chambers were immersed in 10% FBS medium and allowed to migrate for 6 h at 37 °C. Control chambers were inserted in serum-free medium. Cell nuclei were stained with DAPI (0.7 ng/μl) and cell migration was measured by automated cell counting. e-f HUVECs (1 × 10 5 cells/well) were treated with Dis Ba -01 (1000 nM) and/or VEGF (10 ng/mL) and were immediately incubated (37 °C, 1 h) in fibronectin and vitronectin precoated-wells. Negative control was comprised of wells coated with 2% BSA. Cell nuclei were stained with DAPI (0.7 ng/μl) and quantification of adhesion cells was measured by automated cell counting. Results represent the average of three independent experiments in triplicate. Values of * p

    Techniques Used: Migration, Incubation, Spectrophotometry, MTT Assay, Negative Control, Staining, Cell Counting

    Dis Ba -01 inhibits HUVEC tubulogenesis. HUVECs (3 × 10 4 cells/well) were treated for 30 min with VEGF (10 ng/mL), Dis Ba -01 (1, 10, 100 and 1000 nM) or VEGF plus Dis Ba -01 (1000 nM) in DMEM containing 0.5% FBS and then seeded on a solidified Matrigel. The plate was placed in a humidified CO 2 incubator at 37 °C for 14 h to allow the formation of tubes. a Photos (40x magnification) were obtained from a representative experiment ( n = 3). The results were expressed as b Total length (μm 2 ), c Number of mashes, d Number of nodes, e Number of master junctions and f Angiogenesis Score (analysed area x tube length x total of branches). Images were photographed using the AxionVision Rel.4.8 software of a Vert.A1 microscope (Zeiss) and analysed using the Angiogenesis Analyzer plugin for ImageJ software (version 1.51n). Results represent the average of three independent experiments in triplicate. Values of *p
    Figure Legend Snippet: Dis Ba -01 inhibits HUVEC tubulogenesis. HUVECs (3 × 10 4 cells/well) were treated for 30 min with VEGF (10 ng/mL), Dis Ba -01 (1, 10, 100 and 1000 nM) or VEGF plus Dis Ba -01 (1000 nM) in DMEM containing 0.5% FBS and then seeded on a solidified Matrigel. The plate was placed in a humidified CO 2 incubator at 37 °C for 14 h to allow the formation of tubes. a Photos (40x magnification) were obtained from a representative experiment ( n = 3). The results were expressed as b Total length (μm 2 ), c Number of mashes, d Number of nodes, e Number of master junctions and f Angiogenesis Score (analysed area x tube length x total of branches). Images were photographed using the AxionVision Rel.4.8 software of a Vert.A1 microscope (Zeiss) and analysed using the Angiogenesis Analyzer plugin for ImageJ software (version 1.51n). Results represent the average of three independent experiments in triplicate. Values of *p

    Techniques Used: Software, Microscopy

    Dis Ba -01 inhibits VEGFR2 and β 3 phosphorylation after VEGF stimulation. HUVECs (5 × 10 5 cells/well) were seeded in 6-well plates and left to adhere at 37 °C, 5% CO 2 , overnight, followed by a period of 24 h of starvation in serum-free medium. Cells were treated with 1 ml of DMEM supplemented with 10% FBS and Dis Ba -01 (1000 nM), VEGF (10 ng/mL) or a co-treatment and incubated for 1 and 24 h at 37 °C, 5% CO 2 , followed by cell lysis. Twenty micrograms of protein from cell lysates were resolved by SDS-PAGE. Blots were probed with antibodies to a P-TY1054 + TY1059 VEGFR2, to b P-Ty773β 3 and GAPDH, this last to normalize loading. Bands corresponding to all proteins were quantified by densitometry using the ImageJ FIJI program. Bar graph shows the mean ± SE of phosphorylated VEGFR2/GAPDH and β 3 /GAPDH expression from three independent experiments. Values of *p
    Figure Legend Snippet: Dis Ba -01 inhibits VEGFR2 and β 3 phosphorylation after VEGF stimulation. HUVECs (5 × 10 5 cells/well) were seeded in 6-well plates and left to adhere at 37 °C, 5% CO 2 , overnight, followed by a period of 24 h of starvation in serum-free medium. Cells were treated with 1 ml of DMEM supplemented with 10% FBS and Dis Ba -01 (1000 nM), VEGF (10 ng/mL) or a co-treatment and incubated for 1 and 24 h at 37 °C, 5% CO 2 , followed by cell lysis. Twenty micrograms of protein from cell lysates were resolved by SDS-PAGE. Blots were probed with antibodies to a P-TY1054 + TY1059 VEGFR2, to b P-Ty773β 3 and GAPDH, this last to normalize loading. Bands corresponding to all proteins were quantified by densitometry using the ImageJ FIJI program. Bar graph shows the mean ± SE of phosphorylated VEGFR2/GAPDH and β 3 /GAPDH expression from three independent experiments. Values of *p

    Techniques Used: Incubation, Lysis, SDS Page, Expressing

    Dis Ba -01 decreases VEGFR2 protein content. a Analysis of VEGFR2 protein content by western blot. HUVECs (5 × 10 5 cells/well) were seeded in 6-well plates and left to adhere on an incubator at 37 °C, 5% CO 2 , overnight, followed by a period of 24 h of starvation at serum-free medium. Cells were treated with 1 ml of DMEM supplemented with 10% FBS and either Dis Ba -01 (1000 nM), VEGF (10 ng/mL) or a co-treatment and incubated for 1 and 24 h at 37 °C, 5% CO 2 , followed by cell lysis. Twenty micrograms of protein from cell lysates were separated on SDS-PAGE. Blots were probed with VEGFR2 antibody and GAPDH antibody was used to normalize analysis. Bands corresponding to all proteins were quantified by densitometry using the ImageJ FIJI program. Bar graph shows the mean ± SE of VEGFR2/GAPDH expression from three independent experiments. b VEGFR2 mRNA (KDR) expression. HUVECs (5 × 10 5 /well) were seeded in 6-well plates containing DMEM and 10% FBS, followed by a 24-h starvation period on serum-free medium. Cells were treated with Dis Ba -01 (1000 nM) and/or VEGF (10 ng/mL) for 24 h followed by lysis and RNA isolation. Quantitative RT-PCR was carried out using specific primers to human KDR (VEGFR2) and GAPDH (housekeeping). Bar graph shows the mean ± SE of VEGFR2 expression from three independent experiments. Values of *p
    Figure Legend Snippet: Dis Ba -01 decreases VEGFR2 protein content. a Analysis of VEGFR2 protein content by western blot. HUVECs (5 × 10 5 cells/well) were seeded in 6-well plates and left to adhere on an incubator at 37 °C, 5% CO 2 , overnight, followed by a period of 24 h of starvation at serum-free medium. Cells were treated with 1 ml of DMEM supplemented with 10% FBS and either Dis Ba -01 (1000 nM), VEGF (10 ng/mL) or a co-treatment and incubated for 1 and 24 h at 37 °C, 5% CO 2 , followed by cell lysis. Twenty micrograms of protein from cell lysates were separated on SDS-PAGE. Blots were probed with VEGFR2 antibody and GAPDH antibody was used to normalize analysis. Bands corresponding to all proteins were quantified by densitometry using the ImageJ FIJI program. Bar graph shows the mean ± SE of VEGFR2/GAPDH expression from three independent experiments. b VEGFR2 mRNA (KDR) expression. HUVECs (5 × 10 5 /well) were seeded in 6-well plates containing DMEM and 10% FBS, followed by a 24-h starvation period on serum-free medium. Cells were treated with Dis Ba -01 (1000 nM) and/or VEGF (10 ng/mL) for 24 h followed by lysis and RNA isolation. Quantitative RT-PCR was carried out using specific primers to human KDR (VEGFR2) and GAPDH (housekeeping). Bar graph shows the mean ± SE of VEGFR2 expression from three independent experiments. Values of *p

    Techniques Used: Western Blot, Incubation, Lysis, SDS Page, Expressing, Isolation, Quantitative RT-PCR

    4) Product Images from "Recruitment of Stem and Progenitor Cells from the Bone Marrow Niche Requires MMP-9 Mediated Release of Kit-Ligand"

    Article Title: Recruitment of Stem and Progenitor Cells from the Bone Marrow Niche Requires MMP-9 Mediated Release of Kit-Ligand

    Journal: Cell

    doi:

    Chemo/Cytokine-Induced HSC Mobilization Is Impaired in MMP-9 −/− Mice (A–C) MMP-9 −/− and MMP-9 +/+ mice were injected i.v. with a single dose of AdSDF-1, AdVEGF, and AdNull vector or s.c. with recombinant G-CSF from day 0–5 (n = 10 mice in each group). Elevated chemokine levels for SDF-1 and VEGF were achieved by adenoviral gene delivery of SDF-1 and VEGF ([A] and [B], bar graph insert). WBC counts were determined following AdSDF-1 (A), AdVEGF (B), and G-CSF treatment in MMP-9 +/+ mice (C). (D) Mobilized PBMCs were plated in a colony assay. The number of mobilized progenitor cells (CFU-C) was determined (n = 10, *0.05, **p
    Figure Legend Snippet: Chemo/Cytokine-Induced HSC Mobilization Is Impaired in MMP-9 −/− Mice (A–C) MMP-9 −/− and MMP-9 +/+ mice were injected i.v. with a single dose of AdSDF-1, AdVEGF, and AdNull vector or s.c. with recombinant G-CSF from day 0–5 (n = 10 mice in each group). Elevated chemokine levels for SDF-1 and VEGF were achieved by adenoviral gene delivery of SDF-1 and VEGF ([A] and [B], bar graph insert). WBC counts were determined following AdSDF-1 (A), AdVEGF (B), and G-CSF treatment in MMP-9 +/+ mice (C). (D) Mobilized PBMCs were plated in a colony assay. The number of mobilized progenitor cells (CFU-C) was determined (n = 10, *0.05, **p

    Techniques Used: Mouse Assay, Injection, Plasmid Preparation, Recombinant, Colony Assay

    5) Product Images from "Recombinant RGD-disintegrin DisBa-01 blocks integrin αvβ3 and impairs VEGF signaling in endothelial cells"

    Article Title: Recombinant RGD-disintegrin DisBa-01 blocks integrin αvβ3 and impairs VEGF signaling in endothelial cells

    Journal: Cell Communication and Signaling : CCS

    doi: 10.1186/s12964-019-0339-1

    DisBa-01 induces morphological changes in endothelial cells. HUVECs (3 × 10 4 cells/well) were plated in a 96-well microplate previously coated with FN (1 μg/cm 2 ), in serum-free DMEM and incubated overnight at 37 °C, 5% CO 2 . Cells were exposed to VEGF (10 ng/mL), Dis Ba -01 (1000 nM) and VEGF plus Dis Ba -01 for 30 min in DMEM 10% FBS. Cell nuclei were stained with DAPI (0.7 ng/μl) and cytoplasm was stained with Alexa Fluor™ 488 phalloidin for 10 min. Images were observed with 60x magnification. Representative images were obtained from three independent experiments. Scale bar = 50 μm (left panel) and 20 μm (right panel)
    Figure Legend Snippet: DisBa-01 induces morphological changes in endothelial cells. HUVECs (3 × 10 4 cells/well) were plated in a 96-well microplate previously coated with FN (1 μg/cm 2 ), in serum-free DMEM and incubated overnight at 37 °C, 5% CO 2 . Cells were exposed to VEGF (10 ng/mL), Dis Ba -01 (1000 nM) and VEGF plus Dis Ba -01 for 30 min in DMEM 10% FBS. Cell nuclei were stained with DAPI (0.7 ng/μl) and cytoplasm was stained with Alexa Fluor™ 488 phalloidin for 10 min. Images were observed with 60x magnification. Representative images were obtained from three independent experiments. Scale bar = 50 μm (left panel) and 20 μm (right panel)

    Techniques Used: Incubation, Staining

    6) Product Images from "Amino acid restriction triggers angiogenesis via GCN2/ATF4 regulation of VEGF and H2S production"

    Article Title: Amino acid restriction triggers angiogenesis via GCN2/ATF4 regulation of VEGF and H2S production

    Journal: Cell

    doi: 10.1016/j.cell.2018.03.001

    H 2 S promotes glucose uptake and ATP generation by glycolysis for EC migration ( A ) Representative migration across scratch (left, 10X mag) and quantification (right) of HUVEC +/−100μM NaHS in the presence of vehicle or mitomycin C (MitoC, 1μg/mL) to inhibit proliferation; n=12 wells each from cells at 2 different passages; 1-way ANOVA with Sidak’s MCT between control and NaHS within vehicle or MitoC treatment group. ( B, C ) Representative images ( B ) and quantification ( C ) of migration speed (left, n=5–7 cells/condition) and distance (right, n=5–7 cells/condition in x and y directions) from time-lapse video imaging of GFP+ HUVEC infected with control (Ad-Null) or CGL adenovirus (Ad-CGL) as indicated; Student’s T test. ( D ) Relative glucose uptake in HUVEC pretreated with NaHS or 50ng/mL VEGF for 1hr; n=3–6 experiments/group; 1-way ANOVA with Dunnett’s MCT. ( E ) Extracellular acidification rate (ECAR) in WT and CGLKO primary mouse EC pretreated for 1hr with VEGF or NaHS; 10 technical replicates from EC pooled from 6 mice/genotype; 1-way ANOVA with Sidak’s MCT as indicated. ( F ) Glycolytic flux in HUVEC pretreated for 3hr with NaHS or VEGF; 1-way ANOVA with Dunnett’s MCT. Representative experiment of 6 with n=3/group; 1-way ANOVA with Sidak’s MCT. ( G ) Time dependent ATP production in HUVEC pretreated with NaHS or 1mM 2DG at t=0; n=4 experiments each for NaHS and VEGF and 2 for 2DG; error bars indicate SEM; 2-way ANOVA with Dunnett’s MCT relative to t=0 (asterisk, NaHS; caret, VEGF; pound sign, 2DG). ( H, I ) Representative migration (left, 10X mag) and quantification (right, AUC) of HUVEC treated +/−NaHS ( H, n=11 technical replicates/condition) or infected with a control (Ad-Null) or CGL adenovirus (Ad-CGL) at a multiplicity of infection of 50 ( I, n=5–6 technical replicates/condition), in the presence of vehicle or 2DG; 1-way ANOVA with Sidak’s MCT between control and NaHS within 2DG or vehicle treatment group. ( J ) Log2 fold change of C13- labelled metabolites in HUVEC measured by mass spectrometry after 1hr of NaHS pretreatment compared to control; red dots, metabolites with FDR adjusted P
    Figure Legend Snippet: H 2 S promotes glucose uptake and ATP generation by glycolysis for EC migration ( A ) Representative migration across scratch (left, 10X mag) and quantification (right) of HUVEC +/−100μM NaHS in the presence of vehicle or mitomycin C (MitoC, 1μg/mL) to inhibit proliferation; n=12 wells each from cells at 2 different passages; 1-way ANOVA with Sidak’s MCT between control and NaHS within vehicle or MitoC treatment group. ( B, C ) Representative images ( B ) and quantification ( C ) of migration speed (left, n=5–7 cells/condition) and distance (right, n=5–7 cells/condition in x and y directions) from time-lapse video imaging of GFP+ HUVEC infected with control (Ad-Null) or CGL adenovirus (Ad-CGL) as indicated; Student’s T test. ( D ) Relative glucose uptake in HUVEC pretreated with NaHS or 50ng/mL VEGF for 1hr; n=3–6 experiments/group; 1-way ANOVA with Dunnett’s MCT. ( E ) Extracellular acidification rate (ECAR) in WT and CGLKO primary mouse EC pretreated for 1hr with VEGF or NaHS; 10 technical replicates from EC pooled from 6 mice/genotype; 1-way ANOVA with Sidak’s MCT as indicated. ( F ) Glycolytic flux in HUVEC pretreated for 3hr with NaHS or VEGF; 1-way ANOVA with Dunnett’s MCT. Representative experiment of 6 with n=3/group; 1-way ANOVA with Sidak’s MCT. ( G ) Time dependent ATP production in HUVEC pretreated with NaHS or 1mM 2DG at t=0; n=4 experiments each for NaHS and VEGF and 2 for 2DG; error bars indicate SEM; 2-way ANOVA with Dunnett’s MCT relative to t=0 (asterisk, NaHS; caret, VEGF; pound sign, 2DG). ( H, I ) Representative migration (left, 10X mag) and quantification (right, AUC) of HUVEC treated +/−NaHS ( H, n=11 technical replicates/condition) or infected with a control (Ad-Null) or CGL adenovirus (Ad-CGL) at a multiplicity of infection of 50 ( I, n=5–6 technical replicates/condition), in the presence of vehicle or 2DG; 1-way ANOVA with Sidak’s MCT between control and NaHS within 2DG or vehicle treatment group. ( J ) Log2 fold change of C13- labelled metabolites in HUVEC measured by mass spectrometry after 1hr of NaHS pretreatment compared to control; red dots, metabolites with FDR adjusted P

    Techniques Used: Migration, Imaging, Infection, Mouse Assay, Mass Spectrometry

    GCN2-dependent, hypoxia-independent regulation of VEGF and angiogenesis upon SAA restriction ( A) Relative VEGF mRNA expression in HUVEC 2d after transfection with HIF1α siRNA or control scrambled (Sble) siRNA and cultured in control (Ctrl) or SAA deficient (-M C) media for 16hr; n=5 experiments/group; error bars indicate SEM. ( B ) Immunoblots of HIF1α, eIF2α (p- Ser51, total) and ATF4 in HUVEC cultured as indicated for 16hr. ( C ) Relative VEGF mRNA expression in HUVEC 2d after transfection with ATF4 or Sble siRNA and cultured as indicated for 16hr; n=4 experiments/group; SEM. ( D, E ) Relative HUVEC VEGF mRNA expression ( D, n=3 experiments/group; SEM) and secreted VEGF protein concentration in media ( E, n=3–6 experiments/group; SEM) 2d after transfection with ATF4 overexpression (ATF4 OE ) or control construct (Empty). ( F, G ) VEGF mRNA expression ( F ) and spheroid formation ( G ) in WT and GCN2KO primary mouse EC from n=3 mice/genotype cultured as indicated for 16hr. For sprouting assay ( G ), representative images (left, 40X mag) and quantification (right) of WT and GCN2KO EC spheroids cultured in the indicated media for 24hr; blue, DNA (DAPI); red, F-actin (phalloidin). ( H ) Representative transverse sections (left, 40X mag) and quantification (right) of CD31-stained gastroc in WT or GCN2KO mice fed for 2–4wk on Ctrl or MR diets; n=5–6 mice/group. ( I ) VEGF mRNA in MDF, MEF or C2C12 myotubes cultured as indicated for 16hr; n=4–6 experiments/group; SEM. ( J ) VEGF mRNA expression in WT and GCN2KO primary mouse skeletal myotubes (n=5 mice/genotype tested at 2 different passages) cultured as indicated for 16hr. ( K ) Immunoblots of HIF1α, PGC1α, eIF2α (p-Ser51, total) and ATF4 in C2C12 myotubes cultured as indicated for 16hr. Error bars indicate SD unless otherwise noted; asterisks indicate the significance of the difference by Student’s T test or 1-way ANOVA with Sidak’s MCT between diets in vivo or SAA deprivation in vitro ; * P
    Figure Legend Snippet: GCN2-dependent, hypoxia-independent regulation of VEGF and angiogenesis upon SAA restriction ( A) Relative VEGF mRNA expression in HUVEC 2d after transfection with HIF1α siRNA or control scrambled (Sble) siRNA and cultured in control (Ctrl) or SAA deficient (-M C) media for 16hr; n=5 experiments/group; error bars indicate SEM. ( B ) Immunoblots of HIF1α, eIF2α (p- Ser51, total) and ATF4 in HUVEC cultured as indicated for 16hr. ( C ) Relative VEGF mRNA expression in HUVEC 2d after transfection with ATF4 or Sble siRNA and cultured as indicated for 16hr; n=4 experiments/group; SEM. ( D, E ) Relative HUVEC VEGF mRNA expression ( D, n=3 experiments/group; SEM) and secreted VEGF protein concentration in media ( E, n=3–6 experiments/group; SEM) 2d after transfection with ATF4 overexpression (ATF4 OE ) or control construct (Empty). ( F, G ) VEGF mRNA expression ( F ) and spheroid formation ( G ) in WT and GCN2KO primary mouse EC from n=3 mice/genotype cultured as indicated for 16hr. For sprouting assay ( G ), representative images (left, 40X mag) and quantification (right) of WT and GCN2KO EC spheroids cultured in the indicated media for 24hr; blue, DNA (DAPI); red, F-actin (phalloidin). ( H ) Representative transverse sections (left, 40X mag) and quantification (right) of CD31-stained gastroc in WT or GCN2KO mice fed for 2–4wk on Ctrl or MR diets; n=5–6 mice/group. ( I ) VEGF mRNA in MDF, MEF or C2C12 myotubes cultured as indicated for 16hr; n=4–6 experiments/group; SEM. ( J ) VEGF mRNA expression in WT and GCN2KO primary mouse skeletal myotubes (n=5 mice/genotype tested at 2 different passages) cultured as indicated for 16hr. ( K ) Immunoblots of HIF1α, PGC1α, eIF2α (p-Ser51, total) and ATF4 in C2C12 myotubes cultured as indicated for 16hr. Error bars indicate SD unless otherwise noted; asterisks indicate the significance of the difference by Student’s T test or 1-way ANOVA with Sidak’s MCT between diets in vivo or SAA deprivation in vitro ; * P

    Techniques Used: Expressing, Transfection, Cell Culture, Western Blot, Protein Concentration, Over Expression, Construct, Mouse Assay, Staining, In Vivo, In Vitro

    VEGF signalling and AASR converge on endothelial H 2 S production by CGL ( A ) Representative H 2 S production capacity as indicated by black lead sulfide formation from HUVEC cultured in media +/−M C or VEGF (50ng/mL) in the presence or absence of the CGL inhibitor PAG (100μM) as indicated for 16hr. ( B ) Representative (left) endogenous H 2 S levels (blue, H 2 S (P3 fluorescence); red, DNA (DRAQ5)) and quantification of P3 intensity (right) in HUVEC upon VEGF or -M C treatment; n=4 wells/treatment with 4–6 images/well; 1-way ANOVA with Sidak’s MCT vs. Control (asterisks) or +/−PAG within treatment (carets). ( C ) CGL mRNA expression in WT and GCN2KO primary mouse EC cultured from n=3 mice/genotype in control (Ctrl) or -M C media for 16hr. ( D ) CGL mRNA expression in HUVEC 2d after transfection with ATF4 or control scrambled (Sble) siRNA and cultured in the indicated media for 16hr; n=4 experiments/group; SEM. ( E ) CGL mRNA expression in HUVEC 2d after transfection with ATF4 overexpression or control (empty) plasmid; n=3 experiments/group; SEM. ( F, G ) Representative images (left, 40X mag) and quantification (right, in triplicate) of spheroids cultured from ( F ) HUVEC +/−M C for 24hr in the presence of vehicle (Veh) or PAG, and ( G ) WT or CGLKO primary EC sprouts in control or -M C media for 24hr; blue, DNA (DAPI); red, F-actin (phalloidin). Unless otherwise indicated, error bars indicate SD, and asterisks indicate the significance of the difference between diets in vivo or SAA levels in vitro by Student’s T test or 1-way ANOVA with Sidak’s MCT; * P
    Figure Legend Snippet: VEGF signalling and AASR converge on endothelial H 2 S production by CGL ( A ) Representative H 2 S production capacity as indicated by black lead sulfide formation from HUVEC cultured in media +/−M C or VEGF (50ng/mL) in the presence or absence of the CGL inhibitor PAG (100μM) as indicated for 16hr. ( B ) Representative (left) endogenous H 2 S levels (blue, H 2 S (P3 fluorescence); red, DNA (DRAQ5)) and quantification of P3 intensity (right) in HUVEC upon VEGF or -M C treatment; n=4 wells/treatment with 4–6 images/well; 1-way ANOVA with Sidak’s MCT vs. Control (asterisks) or +/−PAG within treatment (carets). ( C ) CGL mRNA expression in WT and GCN2KO primary mouse EC cultured from n=3 mice/genotype in control (Ctrl) or -M C media for 16hr. ( D ) CGL mRNA expression in HUVEC 2d after transfection with ATF4 or control scrambled (Sble) siRNA and cultured in the indicated media for 16hr; n=4 experiments/group; SEM. ( E ) CGL mRNA expression in HUVEC 2d after transfection with ATF4 overexpression or control (empty) plasmid; n=3 experiments/group; SEM. ( F, G ) Representative images (left, 40X mag) and quantification (right, in triplicate) of spheroids cultured from ( F ) HUVEC +/−M C for 24hr in the presence of vehicle (Veh) or PAG, and ( G ) WT or CGLKO primary EC sprouts in control or -M C media for 24hr; blue, DNA (DAPI); red, F-actin (phalloidin). Unless otherwise indicated, error bars indicate SD, and asterisks indicate the significance of the difference between diets in vivo or SAA levels in vitro by Student’s T test or 1-way ANOVA with Sidak’s MCT; * P

    Techniques Used: Cell Culture, Fluorescence, Expressing, Mouse Assay, Transfection, Over Expression, Plasmid Preparation, In Vivo, In Vitro

    SAA restriction induces endothelial VEGF expression in vitro and functional angiogenesis in vivo ( A ) VEGF mRNA levels (left, n=4 experiments/group) and secreted protein concentration in the media (right, n=6 experiments/group) of HUVEC cultured in control (Ctrl) or SAA deficient (- M C) media for 16hr; error bars indicate SEM. (B) Migration assay: Representative migration across a scratch (left, 10X mag at t=20hr; dotted lines indicate boundary of the scratch at t= 0hr) and area under the curve (AUC, right, n=7–10 data points/condition, with each data point representing the mean of multiple measures within a single well in a representative experiment) from HUVEC cultured in the indicated media. ( C ) Tube formation assay: Representative capillary-like structures (left, 40X mag) and quantification of tube length/field in arbitrary units (AU, right; n=8–10 data points/condition) in HUVEC incubated in the indicated media +/−SIRT1 inhibitor Ex-527 for 18hr. ( D ) Spheroid assay: Representative images (left, 40X mag) and quantification (right, in triplicate) of sprouting HUVEC spheroids in the indicated media +/− VEGFR2 inhibitor SU5416 for 24hr; blue, DNA (DAPI); red, F-actin (phalloidin). ( E ) Representative transverse sections (left, 40X mag) and quantification (right) of gastrocnemius muscle stained for endothelial marker CD31 in mice fed 2wk on Ctrl or MR diet +/−VEGFR2 inhibitor axitinib; n=6–8 mice/group. ( F ) Longitudinal Doppler imaging of blood flow in WT mice preconditioned 1mo on Ctrl or MR diet prior to femoral artery ligation (I, ischemic; NI, non-ischemic). Left: representative infrared images on the indicated day after ligation. Right: quantification of blood flow recovery with individual animal AUCs used for statistical comparison; n=7–8 mice/group. ( G ) Representative transverse sections (left, 40X mag) and quantification (right) of CD31-stained gastroc 10d after ligation from ( F ); n=4 mice/group. Error bars indicate SD unless otherwise noted; asterisks indicate the significance of the difference by Student’s T test or 1-way ANOVA with Sidak’s MCT between diets in vivo or SAA deprivation in vitro ; * P
    Figure Legend Snippet: SAA restriction induces endothelial VEGF expression in vitro and functional angiogenesis in vivo ( A ) VEGF mRNA levels (left, n=4 experiments/group) and secreted protein concentration in the media (right, n=6 experiments/group) of HUVEC cultured in control (Ctrl) or SAA deficient (- M C) media for 16hr; error bars indicate SEM. (B) Migration assay: Representative migration across a scratch (left, 10X mag at t=20hr; dotted lines indicate boundary of the scratch at t= 0hr) and area under the curve (AUC, right, n=7–10 data points/condition, with each data point representing the mean of multiple measures within a single well in a representative experiment) from HUVEC cultured in the indicated media. ( C ) Tube formation assay: Representative capillary-like structures (left, 40X mag) and quantification of tube length/field in arbitrary units (AU, right; n=8–10 data points/condition) in HUVEC incubated in the indicated media +/−SIRT1 inhibitor Ex-527 for 18hr. ( D ) Spheroid assay: Representative images (left, 40X mag) and quantification (right, in triplicate) of sprouting HUVEC spheroids in the indicated media +/− VEGFR2 inhibitor SU5416 for 24hr; blue, DNA (DAPI); red, F-actin (phalloidin). ( E ) Representative transverse sections (left, 40X mag) and quantification (right) of gastrocnemius muscle stained for endothelial marker CD31 in mice fed 2wk on Ctrl or MR diet +/−VEGFR2 inhibitor axitinib; n=6–8 mice/group. ( F ) Longitudinal Doppler imaging of blood flow in WT mice preconditioned 1mo on Ctrl or MR diet prior to femoral artery ligation (I, ischemic; NI, non-ischemic). Left: representative infrared images on the indicated day after ligation. Right: quantification of blood flow recovery with individual animal AUCs used for statistical comparison; n=7–8 mice/group. ( G ) Representative transverse sections (left, 40X mag) and quantification (right) of CD31-stained gastroc 10d after ligation from ( F ); n=4 mice/group. Error bars indicate SD unless otherwise noted; asterisks indicate the significance of the difference by Student’s T test or 1-way ANOVA with Sidak’s MCT between diets in vivo or SAA deprivation in vitro ; * P

    Techniques Used: Expressing, In Vitro, Functional Assay, In Vivo, Protein Concentration, Cell Culture, Migration, Tube Formation Assay, Incubation, Staining, Marker, Mouse Assay, Imaging, Flow Cytometry, Ligation

    H 2 S promotes glucose uptake and ATP generation by glycolysis for EC migration ( A ) Representative migration across scratch (left, 10X mag) and quantification (right) of HUVEC +/−100μM NaHS in the presence of vehicle or mitomycin C (MitoC, 1μg/mL) to inhibit proliferation; n=12 wells each from cells at 2 different passages; 1-way ANOVA with Sidak’s MCT between control and NaHS within vehicle or MitoC treatment group. ( B, C ) Representative images ( B ) and quantification ( C ) of migration speed (left, n=5–7 cells/condition) and distance (right, n=5–7 cells/condition in x and y directions) from time-lapse video imaging of GFP+ HUVEC infected with control (Ad-Null) or CGL adenovirus (Ad-CGL) as indicated; Student’s T test. ( D ) Relative glucose uptake in HUVEC pretreated with NaHS or 50ng/mL VEGF for 1hr; n=3–6 experiments/group; 1-way ANOVA with Dunnett’s MCT. ( E ) Extracellular acidification rate (ECAR) in WT and CGLKO primary mouse EC pretreated for 1hr with VEGF or NaHS; 10 technical replicates from EC pooled from 6 mice/genotype; 1-way ANOVA with Sidak’s MCT as indicated. ( F ) Glycolytic flux in HUVEC pretreated for 3hr with NaHS or VEGF; 1-way ANOVA with Dunnett’s MCT. Representative experiment of 6 with n=3/group; 1-way ANOVA with Sidak’s MCT. ( G ) Time dependent ATP production in HUVEC pretreated with NaHS or 1mM 2DG at t=0; n=4 experiments each for NaHS and VEGF and 2 for 2DG; error bars indicate SEM; 2-way ANOVA with Dunnett’s MCT relative to t=0 (asterisk, NaHS; caret, VEGF; pound sign, 2DG). ( H, I ) Representative migration (left, 10X mag) and quantification (right, AUC) of HUVEC treated +/−NaHS ( H, n=11 technical replicates/condition) or infected with a control (Ad-Null) or CGL adenovirus (Ad-CGL) at a multiplicity of infection of 50 ( I, n=5–6 technical replicates/condition), in the presence of vehicle or 2DG; 1-way ANOVA with Sidak’s MCT between control and NaHS within 2DG or vehicle treatment group. ( J ) Log2 fold change of C13- labelled metabolites in HUVEC measured by mass spectrometry after 1hr of NaHS pretreatment compared to control; red dots, metabolites with FDR adjusted P
    Figure Legend Snippet: H 2 S promotes glucose uptake and ATP generation by glycolysis for EC migration ( A ) Representative migration across scratch (left, 10X mag) and quantification (right) of HUVEC +/−100μM NaHS in the presence of vehicle or mitomycin C (MitoC, 1μg/mL) to inhibit proliferation; n=12 wells each from cells at 2 different passages; 1-way ANOVA with Sidak’s MCT between control and NaHS within vehicle or MitoC treatment group. ( B, C ) Representative images ( B ) and quantification ( C ) of migration speed (left, n=5–7 cells/condition) and distance (right, n=5–7 cells/condition in x and y directions) from time-lapse video imaging of GFP+ HUVEC infected with control (Ad-Null) or CGL adenovirus (Ad-CGL) as indicated; Student’s T test. ( D ) Relative glucose uptake in HUVEC pretreated with NaHS or 50ng/mL VEGF for 1hr; n=3–6 experiments/group; 1-way ANOVA with Dunnett’s MCT. ( E ) Extracellular acidification rate (ECAR) in WT and CGLKO primary mouse EC pretreated for 1hr with VEGF or NaHS; 10 technical replicates from EC pooled from 6 mice/genotype; 1-way ANOVA with Sidak’s MCT as indicated. ( F ) Glycolytic flux in HUVEC pretreated for 3hr with NaHS or VEGF; 1-way ANOVA with Dunnett’s MCT. Representative experiment of 6 with n=3/group; 1-way ANOVA with Sidak’s MCT. ( G ) Time dependent ATP production in HUVEC pretreated with NaHS or 1mM 2DG at t=0; n=4 experiments each for NaHS and VEGF and 2 for 2DG; error bars indicate SEM; 2-way ANOVA with Dunnett’s MCT relative to t=0 (asterisk, NaHS; caret, VEGF; pound sign, 2DG). ( H, I ) Representative migration (left, 10X mag) and quantification (right, AUC) of HUVEC treated +/−NaHS ( H, n=11 technical replicates/condition) or infected with a control (Ad-Null) or CGL adenovirus (Ad-CGL) at a multiplicity of infection of 50 ( I, n=5–6 technical replicates/condition), in the presence of vehicle or 2DG; 1-way ANOVA with Sidak’s MCT between control and NaHS within 2DG or vehicle treatment group. ( J ) Log2 fold change of C13- labelled metabolites in HUVEC measured by mass spectrometry after 1hr of NaHS pretreatment compared to control; red dots, metabolites with FDR adjusted P

    Techniques Used: Migration, Imaging, Infection, Mouse Assay, Mass Spectrometry

    7) Product Images from "Inhibition of angiogenesis by arsenic trioxide via TSP-1–TGF-β1-CTGF–VEGF functional module in rheumatoid arthritis"

    Article Title: Inhibition of angiogenesis by arsenic trioxide via TSP-1–TGF-β1-CTGF–VEGF functional module in rheumatoid arthritis

    Journal: Oncotarget

    doi: 10.18632/oncotarget.19867

    The modulation of TSP-1, TGF-β1, CTGF and VEGF expression alongside interventions of TSP-1, TGF-β1, CTGF and VEGF stimulation or knockdown
    Figure Legend Snippet: The modulation of TSP-1, TGF-β1, CTGF and VEGF expression alongside interventions of TSP-1, TGF-β1, CTGF and VEGF stimulation or knockdown

    Techniques Used: Expressing

    As 2 O 3 inhibited angiogenesis by modulating TSP-1, TGF-β1, CTGF and VEGF expression in RA-FLS
    Figure Legend Snippet: As 2 O 3 inhibited angiogenesis by modulating TSP-1, TGF-β1, CTGF and VEGF expression in RA-FLS

    Techniques Used: Expressing

    As 2 O 3 suppressed TSP-1, TGF-β1, CTGF and VEGF expression and microvessel density in synovial tissue of CIA mice
    Figure Legend Snippet: As 2 O 3 suppressed TSP-1, TGF-β1, CTGF and VEGF expression and microvessel density in synovial tissue of CIA mice

    Techniques Used: Expressing, Mouse Assay

    Increased expression of TSP-1, TGF-β1, CTGF and VEGF in supernatants of RA-FLS and HDMECs co-culture compared to NH-FLS and HDMECs co-culture
    Figure Legend Snippet: Increased expression of TSP-1, TGF-β1, CTGF and VEGF in supernatants of RA-FLS and HDMECs co-culture compared to NH-FLS and HDMECs co-culture

    Techniques Used: Expressing, Co-Culture Assay

    8) Product Images from "Effect of radiation-induced endothelial cell injury on platelet regeneration by megakaryocytes"

    Article Title: Effect of radiation-induced endothelial cell injury on platelet regeneration by megakaryocytes

    Journal: Journal of Radiation Research

    doi: 10.1093/jrr/rrx015

    Decreased expression and secretion of VEGF in HUVECs after radiation. HUVEC monolayers were irradiated with 5 Gy radiation and incubated at 37°C for 96 h. (A) The mRNA levels of VEGF were analyzed by qRT-PCR. ** P
    Figure Legend Snippet: Decreased expression and secretion of VEGF in HUVECs after radiation. HUVEC monolayers were irradiated with 5 Gy radiation and incubated at 37°C for 96 h. (A) The mRNA levels of VEGF were analyzed by qRT-PCR. ** P

    Techniques Used: Expressing, Irradiation, Incubation, Quantitative RT-PCR

    9) Product Images from "Cell-surface perturbations of the epidermal growth factor and vascular endothelial growth factor receptors by phosphorothioate oligodeoxynucleotides"

    Article Title: Cell-surface perturbations of the epidermal growth factor and vascular endothelial growth factor receptors by phosphorothioate oligodeoxynucleotides

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi:

    ( A ) Effect of phosphorothioate oligodeoxynucleotides on VEGF-induced activation of the flk-1/fms receptor. Each lane represents the phosphorylated receptor immunoprecipitated from an equal number of C441 cells prebound with 2 μM oligodeoxynucleotide and stimulated with 20 ng/ml VEGF for 15 min at room temperature. Cells were either washed with cold PBS (lanes 1–4) or unwashed (lanes 5–8) prior to the addition of ligand. The level of flk-1/fms activation induced by VEGF is shown for cells preincubated with 2 μM of the oligodeoxynucleotides SdC28 (lanes 1 and 5), #2 (lanes 2 and 6), or without oligodeoxynucleotide (lanes 3 and 7) relative to unstimulated controls (lanes 4 and 8). ( B ) A densitometry scan of A . Data plotted showing flk-1/fms phosphorylation levels with the measurement for the unstimulated control subtracted. ( C ) A reprobing of the blot with an anti-fms polyclonal antibody showing the same level of receptor in each lane.
    Figure Legend Snippet: ( A ) Effect of phosphorothioate oligodeoxynucleotides on VEGF-induced activation of the flk-1/fms receptor. Each lane represents the phosphorylated receptor immunoprecipitated from an equal number of C441 cells prebound with 2 μM oligodeoxynucleotide and stimulated with 20 ng/ml VEGF for 15 min at room temperature. Cells were either washed with cold PBS (lanes 1–4) or unwashed (lanes 5–8) prior to the addition of ligand. The level of flk-1/fms activation induced by VEGF is shown for cells preincubated with 2 μM of the oligodeoxynucleotides SdC28 (lanes 1 and 5), #2 (lanes 2 and 6), or without oligodeoxynucleotide (lanes 3 and 7) relative to unstimulated controls (lanes 4 and 8). ( B ) A densitometry scan of A . Data plotted showing flk-1/fms phosphorylation levels with the measurement for the unstimulated control subtracted. ( C ) A reprobing of the blot with an anti-fms polyclonal antibody showing the same level of receptor in each lane.

    Techniques Used: Activation Assay, Immunoprecipitation

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    Expressing:

    Article Title: Hypoxia Mediates Runt-Related Transcription Factor 2 Expression via Induction of Vascular Endothelial Growth Factor in Periodontal Ligament Stem Cells
    Article Snippet: For RUNX2, basal mRNA expression was decreased by YC-1 in comparison with the normal condition , indicating that HIF-1α was involved in the basal mRNA expression of RUNX2. .. VEGF promotes RUNX2 expression Exogenous recombinant human VEGF (hVEGF165 ) (Peprotech, USA) at different concentrations (0, 1, 2.5, 5, and 10 nmol/L) was added to PDLSCs, and Western blotting showed a concentration-dependent increase in RUNX2 after adding hVEGF165 . ..

    Western Blot:

    Article Title: Hypoxia Mediates Runt-Related Transcription Factor 2 Expression via Induction of Vascular Endothelial Growth Factor in Periodontal Ligament Stem Cells
    Article Snippet: For RUNX2, basal mRNA expression was decreased by YC-1 in comparison with the normal condition , indicating that HIF-1α was involved in the basal mRNA expression of RUNX2. .. VEGF promotes RUNX2 expression Exogenous recombinant human VEGF (hVEGF165 ) (Peprotech, USA) at different concentrations (0, 1, 2.5, 5, and 10 nmol/L) was added to PDLSCs, and Western blotting showed a concentration-dependent increase in RUNX2 after adding hVEGF165 . ..

    Concentration Assay:

    Article Title: Hypoxia Mediates Runt-Related Transcription Factor 2 Expression via Induction of Vascular Endothelial Growth Factor in Periodontal Ligament Stem Cells
    Article Snippet: For RUNX2, basal mRNA expression was decreased by YC-1 in comparison with the normal condition , indicating that HIF-1α was involved in the basal mRNA expression of RUNX2. .. VEGF promotes RUNX2 expression Exogenous recombinant human VEGF (hVEGF165 ) (Peprotech, USA) at different concentrations (0, 1, 2.5, 5, and 10 nmol/L) was added to PDLSCs, and Western blotting showed a concentration-dependent increase in RUNX2 after adding hVEGF165 . ..

    Incubation:

    Article Title: Inhibition of lymphangiogenesis in vitro and in vivo by the multikinase inhibitor nintedanib
    Article Snippet: .. After incubation for 24 h, cells were treated with 50 ng/mL recombinant VEGF-C (PeproTech, Rocky Hill, NJ, USA), PDGF-BB (PeproTech), or bFGF (PeproTech) in the presence or absence of nintedanib at concentrations of 0.01, 0.1, and 1 μM. ..

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    PeproTech vegf a165
    Schematic outline of the contribution of ligand binding to HS and NRP1 in endothelial cell responses. (A) Outline of symbols used in panel B. (B) Depiction of <t>VEGF/VEGF</t> receptor complexes induced by <t>VEGF-A165,</t> VEGF-A121, and VEGF-E-NZ2. Note that we do
    Vegf A165, supplied by PeproTech, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vegf a165/product/PeproTech
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    vegf a165 - by Bioz Stars, 2021-04
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    PeproTech vegf
    Elevated expression of phospho-VE-cadherin in aggressive melanoma cells: a VE-cadherin and Y658 VE-cadherin expression in aggressive melanoma cells (MUM 2B, C8161) and poorly melanoma cells (MUM 2C, C81-61), ( b ) <t>pervanadate</t> treatment in MUM 2B and C8161 showed an increase in Y658 expression levels with decrease in VE-cadherin expression, ( c ) silencing VE-cadherin proves that Y658 VE-cadherin antibody is specific in this melanoma models, ( d ) <t>VEGF</t> (80 ng/ml) for 30 min treatment with or without PF-271 1 μM or siFAK for 24 h or 48 h showed a total decrease in the expression of Y658 VE-cadherin
    Vegf, supplied by PeproTech, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vegf/product/PeproTech
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    vegf - by Bioz Stars, 2021-04
    97/100 stars
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    Rabbit Biotinylated Anti Rat VEGF Source Polyclonal Rabbit Formulation Lyophilized Produced from sera of rabbits pre immunized with highly pure 98 recombinant Rat VEGF Rat Vascular Endothelial Growth Factor Anti
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    Goat Biotinylated Anti Human EG VEGF Source Polyclonal Goat Formulation Lyophilized Produced from sera of goats pre immunized with highly pure 98 recombinant hEG VEGF Anti Human EG VEGF specific
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    Image Search Results


    Schematic outline of the contribution of ligand binding to HS and NRP1 in endothelial cell responses. (A) Outline of symbols used in panel B. (B) Depiction of VEGF/VEGF receptor complexes induced by VEGF-A165, VEGF-A121, and VEGF-E-NZ2. Note that we do

    Journal: Blood

    Article Title: Neuropilin-1 in regulation of VEGF-induced activation of p38MAPK and endothelial cell organization

    doi: 10.1182/blood-2007-12-125856

    Figure Lengend Snippet: Schematic outline of the contribution of ligand binding to HS and NRP1 in endothelial cell responses. (A) Outline of symbols used in panel B. (B) Depiction of VEGF/VEGF receptor complexes induced by VEGF-A165, VEGF-A121, and VEGF-E-NZ2. Note that we do

    Article Snippet: The extent of branching was higher for VEGF-A165, but there was a clear and reproducible effect of VEGF-E-NZ2 on endothelial cell organization.

    Techniques: Ligand Binding Assay

    Sprouting angiogenesis and vessel formation in response to the different VEGF ligands. (A) Sprouting of EBs in 3D collagen cultures was induced in response to VEGF-A165 and VEGF-E-NZ2 (arrows) but not to VEGF-A121 or vehicle, as visualized by anti-CD31

    Journal: Blood

    Article Title: Neuropilin-1 in regulation of VEGF-induced activation of p38MAPK and endothelial cell organization

    doi: 10.1182/blood-2007-12-125856

    Figure Lengend Snippet: Sprouting angiogenesis and vessel formation in response to the different VEGF ligands. (A) Sprouting of EBs in 3D collagen cultures was induced in response to VEGF-A165 and VEGF-E-NZ2 (arrows) but not to VEGF-A121 or vehicle, as visualized by anti-CD31

    Article Snippet: The extent of branching was higher for VEGF-A165, but there was a clear and reproducible effect of VEGF-E-NZ2 on endothelial cell organization.

    Techniques:

    VEGF-A121 fails to rescue ISV sprouting and caudal plexus remodeling defect in vegfa knockdown zebrafish. (A) Vegfa MO (9 ng) alone alternatively vegfa MO (9 ng) combined with vegfa mRNA encoding VEGF-A165, VEGF-A121 (100 pg each), or VEGF-E-NZ2 (200

    Journal: Blood

    Article Title: Neuropilin-1 in regulation of VEGF-induced activation of p38MAPK and endothelial cell organization

    doi: 10.1182/blood-2007-12-125856

    Figure Lengend Snippet: VEGF-A121 fails to rescue ISV sprouting and caudal plexus remodeling defect in vegfa knockdown zebrafish. (A) Vegfa MO (9 ng) alone alternatively vegfa MO (9 ng) combined with vegfa mRNA encoding VEGF-A165, VEGF-A121 (100 pg each), or VEGF-E-NZ2 (200

    Article Snippet: The extent of branching was higher for VEGF-A165, but there was a clear and reproducible effect of VEGF-E-NZ2 on endothelial cell organization.

    Techniques:

    p38MAPK induction by VEGF-A165 and VEGF-E-NZ2, but not by VEGF-A121. (A) PAE/VEGFR-2, NRP1 cells were incubated for different time periods with 2 nM of VEGF-A165, VEGF-A121, or VEGF-E-NZ2, followed by analyses, as indicated, for induction of p38MAPK,

    Journal: Blood

    Article Title: Neuropilin-1 in regulation of VEGF-induced activation of p38MAPK and endothelial cell organization

    doi: 10.1182/blood-2007-12-125856

    Figure Lengend Snippet: p38MAPK induction by VEGF-A165 and VEGF-E-NZ2, but not by VEGF-A121. (A) PAE/VEGFR-2, NRP1 cells were incubated for different time periods with 2 nM of VEGF-A165, VEGF-A121, or VEGF-E-NZ2, followed by analyses, as indicated, for induction of p38MAPK,

    Article Snippet: The extent of branching was higher for VEGF-A165, but there was a clear and reproducible effect of VEGF-E-NZ2 on endothelial cell organization.

    Techniques: Incubation

    VEGF-A165, VEGF-A121, and VEGF-E-NZ2 interaction with VEGFR-2, HS, and NRP1. (A) Schematic outline of VEGF ligands used in this study with VEGF-A exon structure and bindings sites for VEGFR-2, HS, and NRP1 indicated. VEGF-A165 and VEGF-A121 were purchased

    Journal: Blood

    Article Title: Neuropilin-1 in regulation of VEGF-induced activation of p38MAPK and endothelial cell organization

    doi: 10.1182/blood-2007-12-125856

    Figure Lengend Snippet: VEGF-A165, VEGF-A121, and VEGF-E-NZ2 interaction with VEGFR-2, HS, and NRP1. (A) Schematic outline of VEGF ligands used in this study with VEGF-A exon structure and bindings sites for VEGFR-2, HS, and NRP1 indicated. VEGF-A165 and VEGF-A121 were purchased

    Article Snippet: The extent of branching was higher for VEGF-A165, but there was a clear and reproducible effect of VEGF-E-NZ2 on endothelial cell organization.

    Techniques:

    Elevated expression of phospho-VE-cadherin in aggressive melanoma cells: a VE-cadherin and Y658 VE-cadherin expression in aggressive melanoma cells (MUM 2B, C8161) and poorly melanoma cells (MUM 2C, C81-61), ( b ) pervanadate treatment in MUM 2B and C8161 showed an increase in Y658 expression levels with decrease in VE-cadherin expression, ( c ) silencing VE-cadherin proves that Y658 VE-cadherin antibody is specific in this melanoma models, ( d ) VEGF (80 ng/ml) for 30 min treatment with or without PF-271 1 μM or siFAK for 24 h or 48 h showed a total decrease in the expression of Y658 VE-cadherin

    Journal: Cell Death and Differentiation

    Article Title: VE-cadherin promotes vasculogenic mimicry by modulating kaiso-dependent gene expression

    doi: 10.1038/s41418-018-0125-4

    Figure Lengend Snippet: Elevated expression of phospho-VE-cadherin in aggressive melanoma cells: a VE-cadherin and Y658 VE-cadherin expression in aggressive melanoma cells (MUM 2B, C8161) and poorly melanoma cells (MUM 2C, C81-61), ( b ) pervanadate treatment in MUM 2B and C8161 showed an increase in Y658 expression levels with decrease in VE-cadherin expression, ( c ) silencing VE-cadherin proves that Y658 VE-cadherin antibody is specific in this melanoma models, ( d ) VEGF (80 ng/ml) for 30 min treatment with or without PF-271 1 μM or siFAK for 24 h or 48 h showed a total decrease in the expression of Y658 VE-cadherin

    Article Snippet: The following reagents were used: PF-562271 (PF-271, Abcam), VEGF (Peprotech), sodium pervanadate V2O5 + 6 NaOH → 2 Na3VO4 + 3 H2O (5), Corning Matrigel Basement Membrane Matrix for in vitro angiogenesis experiments.

    Techniques: Expressing