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  • 94
    Millipore human vegf
    a SR proteins involved in the control of the <t>VEGF</t> splicing pattern. Most SR proteins were up-regulated in acidic conditions both after 6 and 8 h of exposure. b Expression of the SR protein SRp20, in <t>RL95,</t> increased significantly ( p
    Human Vegf, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 69 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    R&D Systems recombinant vascular permeability factor vegf
    a SR proteins involved in the control of the <t>VEGF</t> splicing pattern. Most SR proteins were up-regulated in acidic conditions both after 6 and 8 h of exposure. b Expression of the SR protein SRp20, in <t>RL95,</t> increased significantly ( p
    Recombinant Vascular Permeability Factor Vegf, supplied by R&D Systems, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    ABclonal vegf
    MVD was correlated positively with <t>HIF-1α</t> and <t>VEGF</t> in the normal pregnancy group ( a , b ). MVD was also correlated positively with HIF-1α and VEGF in the missed abortion group ( c , d )
    Vegf, supplied by ABclonal, used in various techniques. Bioz Stars score: 93/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore vegf
    Deletion of TXNIP does not alter <t>VEGFR2/Akt</t> activation under hyperoxia. Wild type (WT) and TXNIP knockout (TKO) mice were subjected to hyperoxia (75% O2, p7–p12). Activation of VEGFR2 ( A ) and Akt ( B ) were examined as downstream signal of <t>VEGF</t> in p12 WT and TKO retinas. TKO showed significant decrease in phosphorylation of VEGFR-2 and Akt compared to WT under normoxic condition. We did not detect significant change in the activation of VEGFR2 and its downstream Akt in retinas from TKO and WT in response to hyperoxia. (*P
    Vegf, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 1007 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Reliatech vegf a
    Crosstalk of HSPC with Apln + ECs in BM Vascular Regeneration and Hematopoietic Reconstitution (A) Diagram depicting the transplantation of wild-type Lin – cells into Ctrl and Vegfr2 iΔApln host mice. (B) Bone vessels at 2.5 weeks after transplantation in control or Vegfr2 iΔApln host mice. Quantification of Emcn + area, percentage of LSK cells, number of BMNCs, B220 + , CD11b + , and CD8 + cells in control (n = 10–11) and Vegfr2 iΔApln (n = 11) host mice. (C) Scheme depicting <t>VEGF-A</t> treatment in combination with transplantation of wild-type Lin – cells. (D) Survival curve of irradiated mice transplanted with 10 4 Lin – cells and intravenous injection of PBS (vehicle; n = 35) or recombinant VEGF-A (n = 20). (E) Bone vessels at 3 weeks after irradiation and treatment with vehicle (PBS + Lin – cells) or VEGF-A (VEGF-A + Lin – cells). Quantification of Emcn + area, percentage of LSK cells, number of BMNCs, B220 + , CD11b + , and CD8 + cells in vehicle (n = 12–14) and VEGF-A (n = 11)-treated mice. (F) Secondary long-term competitive repopulating assay with donor-derived CD45.2 cells from 1 st transplant of vehicle or VEGF-A-treated recipients. Graphs show percentage of CD45.2 donor-derived myeloid cells, B cells, and T cells (vehicle = 5, VEGF-A = 7). (G) Confocal tile scan overview and high-magnification images of GFP and Emcn signals in the Apln-mTmG diaphysis after infusion of vehicle or VEGF-A. Quantification of GFP + cell relative to Emcn + (vehicle = 3, VEGF-A = 3). (H) Apln transcripts in cultured bEnd.3 cells. Actb was used as control (vehicle = 11, VEGF-A = 11). (I) Quantification of CFU-GM number, CD45% and CD11b% in methylcellulose assays (700 Lin – HSPC were seeded). Vehicle (n = 8) or VEGF-A (4 μg/ml; n = 8) were added to standard culture medium. Error bars, mean ± SEM. p values, two-tailed unpaired Student’s t test. See also Figure S7 .
    Vegf A, supplied by Reliatech, used in various techniques. Bioz Stars score: 92/100, based on 46 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Santa Cruz Biotechnology vegf
    Effects of RS on cellular protein levels of <t>VEGF</t> and <t>EGFR</t> Western blot analyses to determine cellular levels of VEGF and EGFR were conducted using samples from all 4 HNSCC cell lines. Protein levels were determined by densitometry and represented relative
    Vegf, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 97/100, based on 4108 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Shenandoah Biotechnology vegf a
    MSC-conditioned media (CM) from cells differentiated in the 2 <t>kPa+VEGF</t> condition improves paracrine signaling capabilities of <t>MSCs.</t> (A) Top panel: Representative images showing endothelial cell (EC) formation of capillary-like structures on the
    Vegf A, supplied by Shenandoah Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    SLIT2 LTD vegf a
    NCK is required for polarized endothelial cell migration. A, Structural domains of <t>NCK1</t> and 2 indicating VEGFR2 and ROBO1 binding domains and the consensus sequence found in proteins that bind NCK. B, Co-immunoprecipitation of ROBO1 and VEGFR2 with NCK1. Cells were treated with PBS, Slit2 (6 nM) or VEGF (3 nM) for 5min followed by NCK1 IP and immunoblot with anti-ROBO1 or anti-VEGFR2 antibodies. C, Combined NCK1/2 knockdown decreases <t>VEGF-A</t> and Slit2-induced sprouting in 3D-fibrin gels. D, Quantification of sprouting (n= 6). Results are presented as mean ± s.e.m., statistical analyses were performed using Mann-Whitney U test. *P
    Vegf A, supplied by SLIT2 LTD, used in various techniques. Bioz Stars score: 92/100, based on 105 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Genentech vegf
    NCK is required for polarized endothelial cell migration. A, Structural domains of <t>NCK1</t> and 2 indicating VEGFR2 and ROBO1 binding domains and the consensus sequence found in proteins that bind NCK. B, Co-immunoprecipitation of ROBO1 and VEGFR2 with NCK1. Cells were treated with PBS, Slit2 (6 nM) or VEGF (3 nM) for 5min followed by NCK1 IP and immunoblot with anti-ROBO1 or anti-VEGFR2 antibodies. C, Combined NCK1/2 knockdown decreases <t>VEGF-A</t> and Slit2-induced sprouting in 3D-fibrin gels. D, Quantification of sprouting (n= 6). Results are presented as mean ± s.e.m., statistical analyses were performed using Mann-Whitney U test. *P
    Vegf, supplied by Genentech, used in various techniques. Bioz Stars score: 92/100, based on 130 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    OriGene vegf a
    NCK is required for polarized endothelial cell migration. A, Structural domains of <t>NCK1</t> and 2 indicating VEGFR2 and ROBO1 binding domains and the consensus sequence found in proteins that bind NCK. B, Co-immunoprecipitation of ROBO1 and VEGFR2 with NCK1. Cells were treated with PBS, Slit2 (6 nM) or VEGF (3 nM) for 5min followed by NCK1 IP and immunoblot with anti-ROBO1 or anti-VEGFR2 antibodies. C, Combined NCK1/2 knockdown decreases <t>VEGF-A</t> and Slit2-induced sprouting in 3D-fibrin gels. D, Quantification of sprouting (n= 6). Results are presented as mean ± s.e.m., statistical analyses were performed using Mann-Whitney U test. *P
    Vegf A, supplied by OriGene, used in various techniques. Bioz Stars score: 93/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Allergan vegf a
    NCK is required for polarized endothelial cell migration. A, Structural domains of <t>NCK1</t> and 2 indicating VEGFR2 and ROBO1 binding domains and the consensus sequence found in proteins that bind NCK. B, Co-immunoprecipitation of ROBO1 and VEGFR2 with NCK1. Cells were treated with PBS, Slit2 (6 nM) or VEGF (3 nM) for 5min followed by NCK1 IP and immunoblot with anti-ROBO1 or anti-VEGFR2 antibodies. C, Combined NCK1/2 knockdown decreases <t>VEGF-A</t> and Slit2-induced sprouting in 3D-fibrin gels. D, Quantification of sprouting (n= 6). Results are presented as mean ± s.e.m., statistical analyses were performed using Mann-Whitney U test. *P
    Vegf A, supplied by Allergan, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    BioVision vegf a
    NCK is required for polarized endothelial cell migration. A, Structural domains of <t>NCK1</t> and 2 indicating VEGFR2 and ROBO1 binding domains and the consensus sequence found in proteins that bind NCK. B, Co-immunoprecipitation of ROBO1 and VEGFR2 with NCK1. Cells were treated with PBS, Slit2 (6 nM) or VEGF (3 nM) for 5min followed by NCK1 IP and immunoblot with anti-ROBO1 or anti-VEGFR2 antibodies. C, Combined NCK1/2 knockdown decreases <t>VEGF-A</t> and Slit2-induced sprouting in 3D-fibrin gels. D, Quantification of sprouting (n= 6). Results are presented as mean ± s.e.m., statistical analyses were performed using Mann-Whitney U test. *P
    Vegf A, supplied by BioVision, used in various techniques. Bioz Stars score: 92/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    GeneTex vegf a
    Conceptual summary of proposed cellular pathways affected by specific NF-κB blockade in the context of inflammation and angiogenesis, based on the current findings. a Endothelial cell (EC) is exposed to inflammatory stimulus TNF-α, which binds to TNFR1 and triggers a signal transduction resulting in phosphorylation of IκBα by IKK2; IκBα is ubiquitinated and degraded; this process enables the nuclear translocation of NF-κB. Upon nuclear translocation, NF-κB up-regulates a variety of pro-inflammatory and pro-angiogenic genes, including TNF-α, CXCL5, CCL2, and HIF-1α. TNF-α has auto- and paracrine effects and activates NF-kB through positive feedback, which amplifies the inflammatory response. CCL2 and CXCL5 have chemotactic effects on monocytes and neutrophils that in turn secrete a variety of cytokines, including TNF-α and <t>VEGF.</t> Nuclear NF-κB also up-regulates HIF-1α and by this mechanism can directly increase VEGF production. VEGF, in turn, acts on the EC, affecting actin polymerization, cytoskeleton composition, cell motility, tube formation and sprouting angiogenesis. b Selective IKK2 inhibition by IMD-0354 inhibits IκBα phosphorylation by IKK2 disrupting NF-κB activation and nuclear translocation. The pro-inflammatory and pro-angiogenic cytokine and chemokine production are substantially diminished. With decreased levels of VEGF, actin cytoskeleton formation, EC motility and migration are all suppressed. The inhibition of NF-kB also reduces the inflammatory response through suppression of the transcription of pro-inflammatory genes
    Vegf A, supplied by GeneTex, used in various techniques. Bioz Stars score: 93/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    PromoCell vegf a
    Conceptual summary of proposed cellular pathways affected by specific NF-κB blockade in the context of inflammation and angiogenesis, based on the current findings. a Endothelial cell (EC) is exposed to inflammatory stimulus TNF-α, which binds to TNFR1 and triggers a signal transduction resulting in phosphorylation of IκBα by IKK2; IκBα is ubiquitinated and degraded; this process enables the nuclear translocation of NF-κB. Upon nuclear translocation, NF-κB up-regulates a variety of pro-inflammatory and pro-angiogenic genes, including TNF-α, CXCL5, CCL2, and HIF-1α. TNF-α has auto- and paracrine effects and activates NF-kB through positive feedback, which amplifies the inflammatory response. CCL2 and CXCL5 have chemotactic effects on monocytes and neutrophils that in turn secrete a variety of cytokines, including TNF-α and <t>VEGF.</t> Nuclear NF-κB also up-regulates HIF-1α and by this mechanism can directly increase VEGF production. VEGF, in turn, acts on the EC, affecting actin polymerization, cytoskeleton composition, cell motility, tube formation and sprouting angiogenesis. b Selective IKK2 inhibition by IMD-0354 inhibits IκBα phosphorylation by IKK2 disrupting NF-κB activation and nuclear translocation. The pro-inflammatory and pro-angiogenic cytokine and chemokine production are substantially diminished. With decreased levels of VEGF, actin cytoskeleton formation, EC motility and migration are all suppressed. The inhibition of NF-kB also reduces the inflammatory response through suppression of the transcription of pro-inflammatory genes
    Vegf A, supplied by PromoCell, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Sino Biological vegf
    Conceptual summary of proposed cellular pathways affected by specific NF-κB blockade in the context of inflammation and angiogenesis, based on the current findings. a Endothelial cell (EC) is exposed to inflammatory stimulus TNF-α, which binds to TNFR1 and triggers a signal transduction resulting in phosphorylation of IκBα by IKK2; IκBα is ubiquitinated and degraded; this process enables the nuclear translocation of NF-κB. Upon nuclear translocation, NF-κB up-regulates a variety of pro-inflammatory and pro-angiogenic genes, including TNF-α, CXCL5, CCL2, and HIF-1α. TNF-α has auto- and paracrine effects and activates NF-kB through positive feedback, which amplifies the inflammatory response. CCL2 and CXCL5 have chemotactic effects on monocytes and neutrophils that in turn secrete a variety of cytokines, including TNF-α and <t>VEGF.</t> Nuclear NF-κB also up-regulates HIF-1α and by this mechanism can directly increase VEGF production. VEGF, in turn, acts on the EC, affecting actin polymerization, cytoskeleton composition, cell motility, tube formation and sprouting angiogenesis. b Selective IKK2 inhibition by IMD-0354 inhibits IκBα phosphorylation by IKK2 disrupting NF-κB activation and nuclear translocation. The pro-inflammatory and pro-angiogenic cytokine and chemokine production are substantially diminished. With decreased levels of VEGF, actin cytoskeleton formation, EC motility and migration are all suppressed. The inhibition of NF-kB also reduces the inflammatory response through suppression of the transcription of pro-inflammatory genes
    Vegf, supplied by Sino Biological, used in various techniques. Bioz Stars score: 93/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    R&D Systems vegf a
    The NOR-1 protector prevents the down-regulation of VCAM-1 promoted by miR-17 and -20. HUVEC were transfected with precursors of either miR-17 or miR-20a (P17 or P20) or a scramble sequence (Scr) in the presence or in the absence of a miScript target protector that blocks the NOR-1 3’-UTR seed sequence recognized by the miR-17 and -20a (NOR1-Prot). Then cells were stimulated with <t>VEGF</t> (100 ng/ml, 2 h). VCAM-1 expression was assessed by real-time PCR (A) and Western-blot (B) . Results are expressed as mean ± SD from at least n = 4. ( p
    Vegf A, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 1412 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Upstate Biotechnology Inc vegf
    miR-329 is downregulated by angiogenic factors resulting in increased CD146 expression. (A and B) HUVECs were cultured in the presence of <t>VEGF</t> (50 ng/ml) or <t>TNF-α</t> (50 ng/ml) or coincubated with NF-κB pathway inhibitor BAY11-7082 (200 ng/ml)
    Vegf, supplied by Upstate Biotechnology Inc, used in various techniques. Bioz Stars score: 92/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Novus Biologicals vegf antibody
    Effect of DHA-supplemented diet on long-term diabetes-induced degenerative changes in rat retina. Retinas isolated 9 months after induction of diabetes were analyzed by quantitative PCR and immunobloting for inflammatory/angiogenic molecule expression. Quantitative PCR analysis of <t>ASM,</t> ICAM-1, <t>VEGF,</t> and IL-1β ( A ) of retinas isolated from rats subjected to a standard diet (control, white bar ; diabetic, black bar ) or a DHA-enriched diet (diabetic, striped bar) is shown. The results are means ± SE from one set of animals, with five to eight animals in each group. * P
    Vegf Antibody, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abnova vegf a
    GNF351-mediated repression of growth factor expression is AHR-dependent in primary FLS. Repression of AHR expression was performed using AHR-specific siRNA in primary FLS from healthy donors (A) or from patients with RA (B). A total of 2 × 10 6 cells were used per gene knockdown sample. The repression of AHR protein expression was confirmed using Western blot analysis after 48 hours of siRNA transfection. Upon confirmation, primary N-FLS transfected with siRNA for 48 hours were pretreated with 500 nM GNF351 for 1 hour, followed by 10 ng/ml IL1B challenge for 4 hours. The levels of EREG , AREG , <t>VEGF-A</t> , and FGF-2 mRNA were determined by quantitative RT-PCR analysis. Data points and bars represent mean ± S.E. of three independent determinations. Data were analyzed using one-way ANOVA followed by Tukey’s multiple-comparisons test. * P
    Vegf A, supplied by Abnova, used in various techniques. Bioz Stars score: 92/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Agilent technologies vegf
    Immunohistochemical staining of high and low expression of COX-2 ( A , D ), <t>EGFR</t> ( B , E ), and <t>VEGF</t> ( C , F ).
    Vegf, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 92/100, based on 186 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    R&D Systems vegf a165 vegf
    Immunohistochemical staining of high and low expression of COX-2 ( A , D ), <t>EGFR</t> ( B , E ), and <t>VEGF</t> ( C , F ).
    Vegf A165 Vegf, supplied by R&D Systems, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    a SR proteins involved in the control of the VEGF splicing pattern. Most SR proteins were up-regulated in acidic conditions both after 6 and 8 h of exposure. b Expression of the SR protein SRp20, in RL95, increased significantly ( p

    Journal: Cancer Microenvironment

    Article Title: Microenvironment Changes (in pH) Affect VEGF Alternative Splicing

    doi: 10.1007/s12307-008-0013-4

    Figure Lengend Snippet: a SR proteins involved in the control of the VEGF splicing pattern. Most SR proteins were up-regulated in acidic conditions both after 6 and 8 h of exposure. b Expression of the SR protein SRp20, in RL95, increased significantly ( p

    Article Snippet: ELISA, Protein Extraction and Western Blotting Culture supernatants from RL95 in different conditions were collected and used to measure human VEGF by ELISA (Oncogene Research Products) under conditions described by the supplier.

    Techniques: Expressing

    VEGF isoforms expression pattern by RL95 cells in response to changes in the microenvironment. By real time RT-PCR ( a ) and ELISA ( b), we can see an increase in VEGF production in acidic and hypoxic (mimicked by CoCl 2 ) conditions. A shift in the VEGF isoforms splicing pattern is more evident at pH 5.5 where VEGF121 expression is significantly different from all other isoforms ( p

    Journal: Cancer Microenvironment

    Article Title: Microenvironment Changes (in pH) Affect VEGF Alternative Splicing

    doi: 10.1007/s12307-008-0013-4

    Figure Lengend Snippet: VEGF isoforms expression pattern by RL95 cells in response to changes in the microenvironment. By real time RT-PCR ( a ) and ELISA ( b), we can see an increase in VEGF production in acidic and hypoxic (mimicked by CoCl 2 ) conditions. A shift in the VEGF isoforms splicing pattern is more evident at pH 5.5 where VEGF121 expression is significantly different from all other isoforms ( p

    Article Snippet: ELISA, Protein Extraction and Western Blotting Culture supernatants from RL95 in different conditions were collected and used to measure human VEGF by ELISA (Oncogene Research Products) under conditions described by the supplier.

    Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    MVD was correlated positively with HIF-1α and VEGF in the normal pregnancy group ( a , b ). MVD was also correlated positively with HIF-1α and VEGF in the missed abortion group ( c , d )

    Journal: Archives of Gynecology and Obstetrics

    Article Title: Early missed abortion is associated with villous angiogenesis via the HIF-1α/VEGF signaling pathway

    doi: 10.1007/s00404-018-4802-9

    Figure Lengend Snippet: MVD was correlated positively with HIF-1α and VEGF in the normal pregnancy group ( a , b ). MVD was also correlated positively with HIF-1α and VEGF in the missed abortion group ( c , d )

    Article Snippet: In accordance with kit instructions (Zhongshan, Beijing, China), rabbit anti-human HIF, VEGF, and CD34 antibodies (dilution 1:200, ABclonal Biotechnology, USA) were used as primary antibodies.

    Techniques:

    Changes in HIF-1α and VEGF mRNA levels in each group as measured by quantitative PCR ( a , b ). Protein expression of HIF-1α and VEGF in differentially treated HTR8/SVneo cells as measured by western blot analyst ( c ). Protein expression relative to GAPDH of HIF-1α ratio ( d ), and VEGF ratio ( e ); Values are means mean ± standard deviation ( n = 3 for each group). Negative control siRNA as NC control. * P

    Journal: Archives of Gynecology and Obstetrics

    Article Title: Early missed abortion is associated with villous angiogenesis via the HIF-1α/VEGF signaling pathway

    doi: 10.1007/s00404-018-4802-9

    Figure Lengend Snippet: Changes in HIF-1α and VEGF mRNA levels in each group as measured by quantitative PCR ( a , b ). Protein expression of HIF-1α and VEGF in differentially treated HTR8/SVneo cells as measured by western blot analyst ( c ). Protein expression relative to GAPDH of HIF-1α ratio ( d ), and VEGF ratio ( e ); Values are means mean ± standard deviation ( n = 3 for each group). Negative control siRNA as NC control. * P

    Article Snippet: In accordance with kit instructions (Zhongshan, Beijing, China), rabbit anti-human HIF, VEGF, and CD34 antibodies (dilution 1:200, ABclonal Biotechnology, USA) were used as primary antibodies.

    Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot, Standard Deviation, Negative Control

    Immunohistochemical staining for MVD, HIF-1α, and VEGF observed in the cytoplasm of syncytiotrophoblast cells and cytotrophoblast cells ( a ). (×400) Tissues from missed abortions showed lower MVD than with normal pregnancy ( b ). Normal pregnancy showed a high expression of HIF-1α and VEGF ( c ). * P

    Journal: Archives of Gynecology and Obstetrics

    Article Title: Early missed abortion is associated with villous angiogenesis via the HIF-1α/VEGF signaling pathway

    doi: 10.1007/s00404-018-4802-9

    Figure Lengend Snippet: Immunohistochemical staining for MVD, HIF-1α, and VEGF observed in the cytoplasm of syncytiotrophoblast cells and cytotrophoblast cells ( a ). (×400) Tissues from missed abortions showed lower MVD than with normal pregnancy ( b ). Normal pregnancy showed a high expression of HIF-1α and VEGF ( c ). * P

    Article Snippet: In accordance with kit instructions (Zhongshan, Beijing, China), rabbit anti-human HIF, VEGF, and CD34 antibodies (dilution 1:200, ABclonal Biotechnology, USA) were used as primary antibodies.

    Techniques: Immunohistochemistry, Staining, Expressing

    Deletion of TXNIP does not alter VEGFR2/Akt activation under hyperoxia. Wild type (WT) and TXNIP knockout (TKO) mice were subjected to hyperoxia (75% O2, p7–p12). Activation of VEGFR2 ( A ) and Akt ( B ) were examined as downstream signal of VEGF in p12 WT and TKO retinas. TKO showed significant decrease in phosphorylation of VEGFR-2 and Akt compared to WT under normoxic condition. We did not detect significant change in the activation of VEGFR2 and its downstream Akt in retinas from TKO and WT in response to hyperoxia. (*P

    Journal: PLoS ONE

    Article Title: Deletion of Thioredoxin Interacting Protein (TXNIP) Augments Hyperoxia-Induced Vaso-Obliteration in a Mouse Model of Oxygen Induced-Retinopathy

    doi: 10.1371/journal.pone.0110388

    Figure Lengend Snippet: Deletion of TXNIP does not alter VEGFR2/Akt activation under hyperoxia. Wild type (WT) and TXNIP knockout (TKO) mice were subjected to hyperoxia (75% O2, p7–p12). Activation of VEGFR2 ( A ) and Akt ( B ) were examined as downstream signal of VEGF in p12 WT and TKO retinas. TKO showed significant decrease in phosphorylation of VEGFR-2 and Akt compared to WT under normoxic condition. We did not detect significant change in the activation of VEGFR2 and its downstream Akt in retinas from TKO and WT in response to hyperoxia. (*P

    Article Snippet: The primary antibodies were purchased as follow: VEGF (Rabbit polyclonal, EMD-Millipore), phosphor-VEGFR2, VEGFR2, phospho-Akt, Akt, phospho-ASK-1, ASK-1, cleaved caspase-3 (Rabbit polyclonal, Cell Signaling Tech, Danvers, MA), total Trx (Mouse monoclonal, Santa Cruz, Dallas, TX), and TXNIP (Rabbit polyclonal, Invitrogen, Grand Island, NY), cleaved PARP (BD Bioscience Pharmingen, San Diego, CA).

    Techniques: Activation Assay, Knock-Out, Mouse Assay

    Representative diagram shows the impact of TXNIP deletion on retina vasculature under both normoxia and hyperoxia. Under normoxia, retinas from TXNIP-deficient mice showed similar VEGF levels, less peroxynitrite (ONOO-) levels, less VEGF receptor-2 (pVEGFR2) activation and upregulated thioredoxin (Trx) that collectively lead to normal vascular development in comparison to WT mice. Under hyperoxia, retinas from WT mice showed higher peroxynitrite formation, less survival Akt activation (pAkt) and upregulated proapoptotic signal of ASK-1 resulting in vaso-obliteration. Retinas from TKO although showed less peroxynitrite levels and maintained Akt activation, retinas experienced significant decreases in thioredoxin (Trx) that shift the balance of the ASK-1-Trx inhibitory complex and increases the activation of the proapoptotic ASK-1 pathway leading to exacerbated vasoobliteration compared to WT.

    Journal: PLoS ONE

    Article Title: Deletion of Thioredoxin Interacting Protein (TXNIP) Augments Hyperoxia-Induced Vaso-Obliteration in a Mouse Model of Oxygen Induced-Retinopathy

    doi: 10.1371/journal.pone.0110388

    Figure Lengend Snippet: Representative diagram shows the impact of TXNIP deletion on retina vasculature under both normoxia and hyperoxia. Under normoxia, retinas from TXNIP-deficient mice showed similar VEGF levels, less peroxynitrite (ONOO-) levels, less VEGF receptor-2 (pVEGFR2) activation and upregulated thioredoxin (Trx) that collectively lead to normal vascular development in comparison to WT mice. Under hyperoxia, retinas from WT mice showed higher peroxynitrite formation, less survival Akt activation (pAkt) and upregulated proapoptotic signal of ASK-1 resulting in vaso-obliteration. Retinas from TKO although showed less peroxynitrite levels and maintained Akt activation, retinas experienced significant decreases in thioredoxin (Trx) that shift the balance of the ASK-1-Trx inhibitory complex and increases the activation of the proapoptotic ASK-1 pathway leading to exacerbated vasoobliteration compared to WT.

    Article Snippet: The primary antibodies were purchased as follow: VEGF (Rabbit polyclonal, EMD-Millipore), phosphor-VEGFR2, VEGFR2, phospho-Akt, Akt, phospho-ASK-1, ASK-1, cleaved caspase-3 (Rabbit polyclonal, Cell Signaling Tech, Danvers, MA), total Trx (Mouse monoclonal, Santa Cruz, Dallas, TX), and TXNIP (Rabbit polyclonal, Invitrogen, Grand Island, NY), cleaved PARP (BD Bioscience Pharmingen, San Diego, CA).

    Techniques: Mouse Assay, Activation Assay

    Crosstalk of HSPC with Apln + ECs in BM Vascular Regeneration and Hematopoietic Reconstitution (A) Diagram depicting the transplantation of wild-type Lin – cells into Ctrl and Vegfr2 iΔApln host mice. (B) Bone vessels at 2.5 weeks after transplantation in control or Vegfr2 iΔApln host mice. Quantification of Emcn + area, percentage of LSK cells, number of BMNCs, B220 + , CD11b + , and CD8 + cells in control (n = 10–11) and Vegfr2 iΔApln (n = 11) host mice. (C) Scheme depicting VEGF-A treatment in combination with transplantation of wild-type Lin – cells. (D) Survival curve of irradiated mice transplanted with 10 4 Lin – cells and intravenous injection of PBS (vehicle; n = 35) or recombinant VEGF-A (n = 20). (E) Bone vessels at 3 weeks after irradiation and treatment with vehicle (PBS + Lin – cells) or VEGF-A (VEGF-A + Lin – cells). Quantification of Emcn + area, percentage of LSK cells, number of BMNCs, B220 + , CD11b + , and CD8 + cells in vehicle (n = 12–14) and VEGF-A (n = 11)-treated mice. (F) Secondary long-term competitive repopulating assay with donor-derived CD45.2 cells from 1 st transplant of vehicle or VEGF-A-treated recipients. Graphs show percentage of CD45.2 donor-derived myeloid cells, B cells, and T cells (vehicle = 5, VEGF-A = 7). (G) Confocal tile scan overview and high-magnification images of GFP and Emcn signals in the Apln-mTmG diaphysis after infusion of vehicle or VEGF-A. Quantification of GFP + cell relative to Emcn + (vehicle = 3, VEGF-A = 3). (H) Apln transcripts in cultured bEnd.3 cells. Actb was used as control (vehicle = 11, VEGF-A = 11). (I) Quantification of CFU-GM number, CD45% and CD11b% in methylcellulose assays (700 Lin – HSPC were seeded). Vehicle (n = 8) or VEGF-A (4 μg/ml; n = 8) were added to standard culture medium. Error bars, mean ± SEM. p values, two-tailed unpaired Student’s t test. See also Figure S7 .

    Journal: Cell Stem Cell

    Article Title: Apelin+ Endothelial Niche Cells Control Hematopoiesis and Mediate Vascular Regeneration after Myeloablative Injury

    doi: 10.1016/j.stem.2019.10.006

    Figure Lengend Snippet: Crosstalk of HSPC with Apln + ECs in BM Vascular Regeneration and Hematopoietic Reconstitution (A) Diagram depicting the transplantation of wild-type Lin – cells into Ctrl and Vegfr2 iΔApln host mice. (B) Bone vessels at 2.5 weeks after transplantation in control or Vegfr2 iΔApln host mice. Quantification of Emcn + area, percentage of LSK cells, number of BMNCs, B220 + , CD11b + , and CD8 + cells in control (n = 10–11) and Vegfr2 iΔApln (n = 11) host mice. (C) Scheme depicting VEGF-A treatment in combination with transplantation of wild-type Lin – cells. (D) Survival curve of irradiated mice transplanted with 10 4 Lin – cells and intravenous injection of PBS (vehicle; n = 35) or recombinant VEGF-A (n = 20). (E) Bone vessels at 3 weeks after irradiation and treatment with vehicle (PBS + Lin – cells) or VEGF-A (VEGF-A + Lin – cells). Quantification of Emcn + area, percentage of LSK cells, number of BMNCs, B220 + , CD11b + , and CD8 + cells in vehicle (n = 12–14) and VEGF-A (n = 11)-treated mice. (F) Secondary long-term competitive repopulating assay with donor-derived CD45.2 cells from 1 st transplant of vehicle or VEGF-A-treated recipients. Graphs show percentage of CD45.2 donor-derived myeloid cells, B cells, and T cells (vehicle = 5, VEGF-A = 7). (G) Confocal tile scan overview and high-magnification images of GFP and Emcn signals in the Apln-mTmG diaphysis after infusion of vehicle or VEGF-A. Quantification of GFP + cell relative to Emcn + (vehicle = 3, VEGF-A = 3). (H) Apln transcripts in cultured bEnd.3 cells. Actb was used as control (vehicle = 11, VEGF-A = 11). (I) Quantification of CFU-GM number, CD45% and CD11b% in methylcellulose assays (700 Lin – HSPC were seeded). Vehicle (n = 8) or VEGF-A (4 μg/ml; n = 8) were added to standard culture medium. Error bars, mean ± SEM. p values, two-tailed unpaired Student’s t test. See also Figure S7 .

    Article Snippet: In VEGF-A treatment experiments, 104 lineage-negative cells were transplanted together with 2 μg VEGF-A (Reliatech, 300-076) 4 to 6 hours after lethal irradiation.

    Techniques: Transplantation Assay, Mouse Assay, Irradiation, Injection, Recombinant, Derivative Assay, Cell Culture, Two Tailed Test

    Effects of RS on cellular protein levels of VEGF and EGFR Western blot analyses to determine cellular levels of VEGF and EGFR were conducted using samples from all 4 HNSCC cell lines. Protein levels were determined by densitometry and represented relative

    Journal:

    Article Title: The Effects of Reactive Species on the Tumorigenic Phenotype of Human Head and Neck Squamous Cell Carcinoma (HNSCC) Cells

    doi:

    Figure Lengend Snippet: Effects of RS on cellular protein levels of VEGF and EGFR Western blot analyses to determine cellular levels of VEGF and EGFR were conducted using samples from all 4 HNSCC cell lines. Protein levels were determined by densitometry and represented relative

    Article Snippet: Samples were run using a 7.5% SDS-PAGE and probed for VEGF (1:800, Santa Cruz Biotechnology), EGFR (1:800, Santa Cruz) and β-actin (1:6666, AB Cam, Cambridge, MA, USA).

    Techniques: Western Blot

    Syncytial streamers (ST) in the subplacenta (SP) and in the decidua expressed VEGF, Flt-1, KDR, B2R and eNOS in days 20, 40 and 60 of pregnancy. Bar = 100 μm. Rectangle defines the area shown at a higher magnification (400×) in Figure 6.

    Journal: Reproductive biology and endocrinology : RB & E

    Article Title: Angiogenic, hyperpermeability and vasodilator network in utero-placental units along pregnancy in the guinea-pig (Cavia porcellus)

    doi: 10.1186/1477-7827-6-13

    Figure Lengend Snippet: Syncytial streamers (ST) in the subplacenta (SP) and in the decidua expressed VEGF, Flt-1, KDR, B2R and eNOS in days 20, 40 and 60 of pregnancy. Bar = 100 μm. Rectangle defines the area shown at a higher magnification (400×) in Figure 6.

    Article Snippet: Sections were incubated in a humid chamber for 30 minutes with protein block (Cas-Block® , Zymed, San Francisco, CA) followed by incubation for 18 h at 4°C with the primary antibodies: mouse monoclonal anti-eNOS (1:50, clone 3 BD Transduction Laboratories), VEGF (1:50, clone JH 121, Upstate); rabbit polyclonal KDR (1:500, Upstate) and Flt-1 (1:1000, Santa Cruz Biotechnology, Inc).

    Techniques:

    Mesometrial arteries obtained in days 20 and 40 of pregnancy showed in sequential sections an intact smooth muscle layer characterized by muscle actin (MA), and no cytokeratin positive cells. In day 60, intramural trophoblasts, characterized by cytokeratin (CK), expressed VEGF, Flt-1, KDR, B2R and eNOS, as did the swollen endothelial cells, identified by von Willebrand Factor (vWF). The remaining vascular smooth muscle, positive for muscle actin (MA), was disrupted. Bar = 100 μm.

    Journal: Reproductive biology and endocrinology : RB & E

    Article Title: Angiogenic, hyperpermeability and vasodilator network in utero-placental units along pregnancy in the guinea-pig (Cavia porcellus)

    doi: 10.1186/1477-7827-6-13

    Figure Lengend Snippet: Mesometrial arteries obtained in days 20 and 40 of pregnancy showed in sequential sections an intact smooth muscle layer characterized by muscle actin (MA), and no cytokeratin positive cells. In day 60, intramural trophoblasts, characterized by cytokeratin (CK), expressed VEGF, Flt-1, KDR, B2R and eNOS, as did the swollen endothelial cells, identified by von Willebrand Factor (vWF). The remaining vascular smooth muscle, positive for muscle actin (MA), was disrupted. Bar = 100 μm.

    Article Snippet: Sections were incubated in a humid chamber for 30 minutes with protein block (Cas-Block® , Zymed, San Francisco, CA) followed by incubation for 18 h at 4°C with the primary antibodies: mouse monoclonal anti-eNOS (1:50, clone 3 BD Transduction Laboratories), VEGF (1:50, clone JH 121, Upstate); rabbit polyclonal KDR (1:500, Upstate) and Flt-1 (1:1000, Santa Cruz Biotechnology, Inc).

    Techniques:

    In subsequent sections of a utero-placental unit obtained in day 15 of pregnancy, the multilayered subplacenta (SP) gave rise to the placental sprouts, and to the syncytial streamers that penetrate the adjacent decidua. The subplacenta, the placental sprouts and the syncytial streamers expressed VEGF, Flt-1, KDR, B2R and eNOS. Cytotrophoblasts were characterized by cytokeratin (CK) staining. Bar = 100 μm.

    Journal: Reproductive biology and endocrinology : RB & E

    Article Title: Angiogenic, hyperpermeability and vasodilator network in utero-placental units along pregnancy in the guinea-pig (Cavia porcellus)

    doi: 10.1186/1477-7827-6-13

    Figure Lengend Snippet: In subsequent sections of a utero-placental unit obtained in day 15 of pregnancy, the multilayered subplacenta (SP) gave rise to the placental sprouts, and to the syncytial streamers that penetrate the adjacent decidua. The subplacenta, the placental sprouts and the syncytial streamers expressed VEGF, Flt-1, KDR, B2R and eNOS. Cytotrophoblasts were characterized by cytokeratin (CK) staining. Bar = 100 μm.

    Article Snippet: Sections were incubated in a humid chamber for 30 minutes with protein block (Cas-Block® , Zymed, San Francisco, CA) followed by incubation for 18 h at 4°C with the primary antibodies: mouse monoclonal anti-eNOS (1:50, clone 3 BD Transduction Laboratories), VEGF (1:50, clone JH 121, Upstate); rabbit polyclonal KDR (1:500, Upstate) and Flt-1 (1:1000, Santa Cruz Biotechnology, Inc).

    Techniques: Staining

    Syncytiotrophoblasts composing the interlobium and the labyrinth expressed Flt-1 and VEGF, in sections obtained in days 20, 40 and 60 of pregnancy at a high magnification (1000×). The immunoreactivity showed a granular pattern for interlobar Flt-1, and for interlobar and labyrinthine VEGF; labyrinthine Flt-1 displayed a diffuse cytoplasmic staining. Arrowheads highlight linear signal in endothelial cells. Bar = 100 μm.

    Journal: Reproductive biology and endocrinology : RB & E

    Article Title: Angiogenic, hyperpermeability and vasodilator network in utero-placental units along pregnancy in the guinea-pig (Cavia porcellus)

    doi: 10.1186/1477-7827-6-13

    Figure Lengend Snippet: Syncytiotrophoblasts composing the interlobium and the labyrinth expressed Flt-1 and VEGF, in sections obtained in days 20, 40 and 60 of pregnancy at a high magnification (1000×). The immunoreactivity showed a granular pattern for interlobar Flt-1, and for interlobar and labyrinthine VEGF; labyrinthine Flt-1 displayed a diffuse cytoplasmic staining. Arrowheads highlight linear signal in endothelial cells. Bar = 100 μm.

    Article Snippet: Sections were incubated in a humid chamber for 30 minutes with protein block (Cas-Block® , Zymed, San Francisco, CA) followed by incubation for 18 h at 4°C with the primary antibodies: mouse monoclonal anti-eNOS (1:50, clone 3 BD Transduction Laboratories), VEGF (1:50, clone JH 121, Upstate); rabbit polyclonal KDR (1:500, Upstate) and Flt-1 (1:1000, Santa Cruz Biotechnology, Inc).

    Techniques: Staining

    Representative blot of placental homogenates from days 20, 40 and 60 of pregnancy for FLT-1 and eNOS. Purified human Flt-1 and eNOS yielded bands with approximate molecular weights of 180 and 140 kDa respectively. No significant differences were observed between the means of the 3 homogenates included in each study period, using one-way analysis of variance and Tukey's Multiple Comparison post-hoc test.

    Journal: Reproductive biology and endocrinology : RB & E

    Article Title: Angiogenic, hyperpermeability and vasodilator network in utero-placental units along pregnancy in the guinea-pig (Cavia porcellus)

    doi: 10.1186/1477-7827-6-13

    Figure Lengend Snippet: Representative blot of placental homogenates from days 20, 40 and 60 of pregnancy for FLT-1 and eNOS. Purified human Flt-1 and eNOS yielded bands with approximate molecular weights of 180 and 140 kDa respectively. No significant differences were observed between the means of the 3 homogenates included in each study period, using one-way analysis of variance and Tukey's Multiple Comparison post-hoc test.

    Article Snippet: Sections were incubated in a humid chamber for 30 minutes with protein block (Cas-Block® , Zymed, San Francisco, CA) followed by incubation for 18 h at 4°C with the primary antibodies: mouse monoclonal anti-eNOS (1:50, clone 3 BD Transduction Laboratories), VEGF (1:50, clone JH 121, Upstate); rabbit polyclonal KDR (1:500, Upstate) and Flt-1 (1:1000, Santa Cruz Biotechnology, Inc).

    Techniques: Purification

    Signaling pathways that contribute to VEGF induced angiogenesis, and a proposal for their participation in the development of the utero-placental interface by participating in proliferative, invasive, vasodilatory and permeability changes essential for cell invasion and angiogenesis. VEGF activates eNOS through pathways including Akt/PKB, Ca +2 /CaM, and PKC [20-24]. Flt-1 may negatively regulate KDR, but might also promote its activity [20]. Bradykinin stimulates eNOS through Ca +2 , induces EC to form tubes, and transactivates the KDR [26]. The factors studied have been depicted in orange areas surrounded by a red border, known mechanisms of downstream activation by interrupted arrows, and unknown mechanism of downstream activation by interrupted arrow. VEGF, vascular endothelial growth factor; BK, bradykinin; eNOS, endothelial nitric oxide synthase; PLC β, phospholipase C -β; PLCγ, phospholipase C -γ; DAG, diacylglycerol; IP3, inositol (1,4,5)-triphosphate; PKC, protein kinase C; MAPK, mitogen-activated protein kinase; FAK, focal adhesion kinase; PI3K, phosphoinositide 3-kinase; p38 MAPK, p38 mitogen-activated protein; Erk, extracellular regulated kinase; HSP27, heat protein; CaM, calmodulin; EC, endothelial cell.

    Journal: Reproductive biology and endocrinology : RB & E

    Article Title: Angiogenic, hyperpermeability and vasodilator network in utero-placental units along pregnancy in the guinea-pig (Cavia porcellus)

    doi: 10.1186/1477-7827-6-13

    Figure Lengend Snippet: Signaling pathways that contribute to VEGF induced angiogenesis, and a proposal for their participation in the development of the utero-placental interface by participating in proliferative, invasive, vasodilatory and permeability changes essential for cell invasion and angiogenesis. VEGF activates eNOS through pathways including Akt/PKB, Ca +2 /CaM, and PKC [20-24]. Flt-1 may negatively regulate KDR, but might also promote its activity [20]. Bradykinin stimulates eNOS through Ca +2 , induces EC to form tubes, and transactivates the KDR [26]. The factors studied have been depicted in orange areas surrounded by a red border, known mechanisms of downstream activation by interrupted arrows, and unknown mechanism of downstream activation by interrupted arrow. VEGF, vascular endothelial growth factor; BK, bradykinin; eNOS, endothelial nitric oxide synthase; PLC β, phospholipase C -β; PLCγ, phospholipase C -γ; DAG, diacylglycerol; IP3, inositol (1,4,5)-triphosphate; PKC, protein kinase C; MAPK, mitogen-activated protein kinase; FAK, focal adhesion kinase; PI3K, phosphoinositide 3-kinase; p38 MAPK, p38 mitogen-activated protein; Erk, extracellular regulated kinase; HSP27, heat protein; CaM, calmodulin; EC, endothelial cell.

    Article Snippet: Sections were incubated in a humid chamber for 30 minutes with protein block (Cas-Block® , Zymed, San Francisco, CA) followed by incubation for 18 h at 4°C with the primary antibodies: mouse monoclonal anti-eNOS (1:50, clone 3 BD Transduction Laboratories), VEGF (1:50, clone JH 121, Upstate); rabbit polyclonal KDR (1:500, Upstate) and Flt-1 (1:1000, Santa Cruz Biotechnology, Inc).

    Techniques: Permeability, Chick Chorioallantoic Membrane Assay, Activity Assay, Activation Assay, Planar Chromatography

    MSC-conditioned media (CM) from cells differentiated in the 2 kPa+VEGF condition improves paracrine signaling capabilities of MSCs. (A) Top panel: Representative images showing endothelial cell (EC) formation of capillary-like structures on the

    Journal: Tissue Engineering. Part A

    Article Title: Synergism of Matrix Stiffness and Vascular Endothelial Growth Factor on Mesenchymal Stem Cells for Vascular Endothelial Regeneration

    doi: 10.1089/ten.tea.2013.0249

    Figure Lengend Snippet: MSC-conditioned media (CM) from cells differentiated in the 2 kPa+VEGF condition improves paracrine signaling capabilities of MSCs. (A) Top panel: Representative images showing endothelial cell (EC) formation of capillary-like structures on the

    Article Snippet: To examine the effect of soluble chemical factors on vascular differentiation of MSCs, VEGF-A (Shenandoah Biotechnology, Warwick, PA) was added to the media.

    Techniques:

    The combination of VEGF and 2 kPa matrix elasticity upregulates genes showing early and mature endothelial phenotype in MSCs. (A) MSC gene expression of Flk-1 , Flt-1 , vWF , and eNOS after 24 and 168 h in seeding conditions. * p

    Journal: Tissue Engineering. Part A

    Article Title: Synergism of Matrix Stiffness and Vascular Endothelial Growth Factor on Mesenchymal Stem Cells for Vascular Endothelial Regeneration

    doi: 10.1089/ten.tea.2013.0249

    Figure Lengend Snippet: The combination of VEGF and 2 kPa matrix elasticity upregulates genes showing early and mature endothelial phenotype in MSCs. (A) MSC gene expression of Flk-1 , Flt-1 , vWF , and eNOS after 24 and 168 h in seeding conditions. * p

    Article Snippet: To examine the effect of soluble chemical factors on vascular differentiation of MSCs, VEGF-A (Shenandoah Biotechnology, Warwick, PA) was added to the media.

    Techniques: Expressing

    Effect of the vascular endothelial growth factor (VEGF) concentration on the mesenchymal stem cell (MSC) expression of early endothelial markers. (A) Representative images of MSC stained with endothelial markers (Flk-1 and VECAD) after 168-h incubation

    Journal: Tissue Engineering. Part A

    Article Title: Synergism of Matrix Stiffness and Vascular Endothelial Growth Factor on Mesenchymal Stem Cells for Vascular Endothelial Regeneration

    doi: 10.1089/ten.tea.2013.0249

    Figure Lengend Snippet: Effect of the vascular endothelial growth factor (VEGF) concentration on the mesenchymal stem cell (MSC) expression of early endothelial markers. (A) Representative images of MSC stained with endothelial markers (Flk-1 and VECAD) after 168-h incubation

    Article Snippet: To examine the effect of soluble chemical factors on vascular differentiation of MSCs, VEGF-A (Shenandoah Biotechnology, Warwick, PA) was added to the media.

    Techniques: Concentration Assay, Expressing, Staining, Incubation

    The combination of VEGF and 2 kPa matrix elasticity upregulates MSC expression of endothelial-specific functional proteins while downregulating SMA expression. (A) Representative immunostaining images and quantitative measures showing SMA + MSCs

    Journal: Tissue Engineering. Part A

    Article Title: Synergism of Matrix Stiffness and Vascular Endothelial Growth Factor on Mesenchymal Stem Cells for Vascular Endothelial Regeneration

    doi: 10.1089/ten.tea.2013.0249

    Figure Lengend Snippet: The combination of VEGF and 2 kPa matrix elasticity upregulates MSC expression of endothelial-specific functional proteins while downregulating SMA expression. (A) Representative immunostaining images and quantitative measures showing SMA + MSCs

    Article Snippet: To examine the effect of soluble chemical factors on vascular differentiation of MSCs, VEGF-A (Shenandoah Biotechnology, Warwick, PA) was added to the media.

    Techniques: Expressing, Functional Assay, Immunostaining

    MSCs formed capillary-like tube structures in 2 kPa and 2 kPa+VEGF seeding conditions. (A) Representative fluorescent (F-actin) and phase contrast images of capillary-like structures formed by MSCs in 2 kPa and 2 kPA+VEGF

    Journal: Tissue Engineering. Part A

    Article Title: Synergism of Matrix Stiffness and Vascular Endothelial Growth Factor on Mesenchymal Stem Cells for Vascular Endothelial Regeneration

    doi: 10.1089/ten.tea.2013.0249

    Figure Lengend Snippet: MSCs formed capillary-like tube structures in 2 kPa and 2 kPa+VEGF seeding conditions. (A) Representative fluorescent (F-actin) and phase contrast images of capillary-like structures formed by MSCs in 2 kPa and 2 kPA+VEGF

    Article Snippet: To examine the effect of soluble chemical factors on vascular differentiation of MSCs, VEGF-A (Shenandoah Biotechnology, Warwick, PA) was added to the media.

    Techniques:

    NCK is required for polarized endothelial cell migration. A, Structural domains of NCK1 and 2 indicating VEGFR2 and ROBO1 binding domains and the consensus sequence found in proteins that bind NCK. B, Co-immunoprecipitation of ROBO1 and VEGFR2 with NCK1. Cells were treated with PBS, Slit2 (6 nM) or VEGF (3 nM) for 5min followed by NCK1 IP and immunoblot with anti-ROBO1 or anti-VEGFR2 antibodies. C, Combined NCK1/2 knockdown decreases VEGF-A and Slit2-induced sprouting in 3D-fibrin gels. D, Quantification of sprouting (n= 6). Results are presented as mean ± s.e.m., statistical analyses were performed using Mann-Whitney U test. *P

    Journal: Circulation

    Article Title: Targeting NCK-Mediated Endothelial Cell Front-Rear Polarity Inhibits Neo-Vascularization

    doi: 10.1161/CIRCULATIONAHA.115.017537

    Figure Lengend Snippet: NCK is required for polarized endothelial cell migration. A, Structural domains of NCK1 and 2 indicating VEGFR2 and ROBO1 binding domains and the consensus sequence found in proteins that bind NCK. B, Co-immunoprecipitation of ROBO1 and VEGFR2 with NCK1. Cells were treated with PBS, Slit2 (6 nM) or VEGF (3 nM) for 5min followed by NCK1 IP and immunoblot with anti-ROBO1 or anti-VEGFR2 antibodies. C, Combined NCK1/2 knockdown decreases VEGF-A and Slit2-induced sprouting in 3D-fibrin gels. D, Quantification of sprouting (n= 6). Results are presented as mean ± s.e.m., statistical analyses were performed using Mann-Whitney U test. *P

    Article Snippet: NCK1/2 coordinates the functional interaction between VEGF-A and Slit2 signaling required for front-rear polarity and sprouting angiogenesis.

    Techniques: Migration, Binding Assay, Sequencing, Immunoprecipitation, MANN-WHITNEY

    Slit2/ROBO1/NCK are required for VEGF-A induced front-rear polarity. A, Retinal IB4 (green), Gm130 (red) and Erg1/2/3 (blue) triple-labeling of mice injected with control or Robo1-Fc adenovirus. B, Quantification of the different Golgi positions around the nuclei toward the migrating front (n=3 retina for Nck1 −/− Nck2 l/l and n=3 retina for Nck1 +/+ Nck2iec, at least 50 tip cells per retina were quantified). Results are presented as mean ± s.e.m., statistical analyses were performed using Mann-Whitney U test. *P

    Journal: Circulation

    Article Title: Targeting NCK-Mediated Endothelial Cell Front-Rear Polarity Inhibits Neo-Vascularization

    doi: 10.1161/CIRCULATIONAHA.115.017537

    Figure Lengend Snippet: Slit2/ROBO1/NCK are required for VEGF-A induced front-rear polarity. A, Retinal IB4 (green), Gm130 (red) and Erg1/2/3 (blue) triple-labeling of mice injected with control or Robo1-Fc adenovirus. B, Quantification of the different Golgi positions around the nuclei toward the migrating front (n=3 retina for Nck1 −/− Nck2 l/l and n=3 retina for Nck1 +/+ Nck2iec, at least 50 tip cells per retina were quantified). Results are presented as mean ± s.e.m., statistical analyses were performed using Mann-Whitney U test. *P

    Article Snippet: NCK1/2 coordinates the functional interaction between VEGF-A and Slit2 signaling required for front-rear polarity and sprouting angiogenesis.

    Techniques: Labeling, Mouse Assay, Injection, MANN-WHITNEY

    Conceptual summary of proposed cellular pathways affected by specific NF-κB blockade in the context of inflammation and angiogenesis, based on the current findings. a Endothelial cell (EC) is exposed to inflammatory stimulus TNF-α, which binds to TNFR1 and triggers a signal transduction resulting in phosphorylation of IκBα by IKK2; IκBα is ubiquitinated and degraded; this process enables the nuclear translocation of NF-κB. Upon nuclear translocation, NF-κB up-regulates a variety of pro-inflammatory and pro-angiogenic genes, including TNF-α, CXCL5, CCL2, and HIF-1α. TNF-α has auto- and paracrine effects and activates NF-kB through positive feedback, which amplifies the inflammatory response. CCL2 and CXCL5 have chemotactic effects on monocytes and neutrophils that in turn secrete a variety of cytokines, including TNF-α and VEGF. Nuclear NF-κB also up-regulates HIF-1α and by this mechanism can directly increase VEGF production. VEGF, in turn, acts on the EC, affecting actin polymerization, cytoskeleton composition, cell motility, tube formation and sprouting angiogenesis. b Selective IKK2 inhibition by IMD-0354 inhibits IκBα phosphorylation by IKK2 disrupting NF-κB activation and nuclear translocation. The pro-inflammatory and pro-angiogenic cytokine and chemokine production are substantially diminished. With decreased levels of VEGF, actin cytoskeleton formation, EC motility and migration are all suppressed. The inhibition of NF-kB also reduces the inflammatory response through suppression of the transcription of pro-inflammatory genes

    Journal: Angiogenesis

    Article Title: Selective IKK2 inhibitor IMD0354 disrupts NF-κB signaling to suppress corneal inflammation and angiogenesis

    doi: 10.1007/s10456-018-9594-9

    Figure Lengend Snippet: Conceptual summary of proposed cellular pathways affected by specific NF-κB blockade in the context of inflammation and angiogenesis, based on the current findings. a Endothelial cell (EC) is exposed to inflammatory stimulus TNF-α, which binds to TNFR1 and triggers a signal transduction resulting in phosphorylation of IκBα by IKK2; IκBα is ubiquitinated and degraded; this process enables the nuclear translocation of NF-κB. Upon nuclear translocation, NF-κB up-regulates a variety of pro-inflammatory and pro-angiogenic genes, including TNF-α, CXCL5, CCL2, and HIF-1α. TNF-α has auto- and paracrine effects and activates NF-kB through positive feedback, which amplifies the inflammatory response. CCL2 and CXCL5 have chemotactic effects on monocytes and neutrophils that in turn secrete a variety of cytokines, including TNF-α and VEGF. Nuclear NF-κB also up-regulates HIF-1α and by this mechanism can directly increase VEGF production. VEGF, in turn, acts on the EC, affecting actin polymerization, cytoskeleton composition, cell motility, tube formation and sprouting angiogenesis. b Selective IKK2 inhibition by IMD-0354 inhibits IκBα phosphorylation by IKK2 disrupting NF-κB activation and nuclear translocation. The pro-inflammatory and pro-angiogenic cytokine and chemokine production are substantially diminished. With decreased levels of VEGF, actin cytoskeleton formation, EC motility and migration are all suppressed. The inhibition of NF-kB also reduces the inflammatory response through suppression of the transcription of pro-inflammatory genes

    Article Snippet: Samples were incubated with VEGF-a (GTX21316, 1:100; GeneTex, Simpson, PA, US) antibody, then visualized (DyLight 519, 1:500; Thermo Fisher Scientific, MA, US), mounted (ProLong Gold antifade regent; Invitrogen, Thermo Fisher Scientific, MA, US) and imaged.

    Techniques: Transduction, Translocation Assay, Inhibition, Radial Immuno Diffusion, Activation Assay, Migration

    Quantitative qRT-PCR and Western blot analysis of inflammatory and neovascular factors in suture-stimulated rat corneas at 96 h. Quantitative qRT-PCR analysis of a Vegf - a , b Cxcl5 , c Ccl2 , and d Cxcr2 expression in rat cornea. ( n = 5) One-way ANOVA test with Tukey multiple comparisons were used to determine statistical significance. e Western blot analysis of CD45, phospho-IκBα and VEGF-A expression in rat cornea lysate. β-Actin used as a loading control. n.s. p > 0.05; * p

    Journal: Angiogenesis

    Article Title: Selective IKK2 inhibitor IMD0354 disrupts NF-κB signaling to suppress corneal inflammation and angiogenesis

    doi: 10.1007/s10456-018-9594-9

    Figure Lengend Snippet: Quantitative qRT-PCR and Western blot analysis of inflammatory and neovascular factors in suture-stimulated rat corneas at 96 h. Quantitative qRT-PCR analysis of a Vegf - a , b Cxcl5 , c Ccl2 , and d Cxcr2 expression in rat cornea. ( n = 5) One-way ANOVA test with Tukey multiple comparisons were used to determine statistical significance. e Western blot analysis of CD45, phospho-IκBα and VEGF-A expression in rat cornea lysate. β-Actin used as a loading control. n.s. p > 0.05; * p

    Article Snippet: Samples were incubated with VEGF-a (GTX21316, 1:100; GeneTex, Simpson, PA, US) antibody, then visualized (DyLight 519, 1:500; Thermo Fisher Scientific, MA, US), mounted (ProLong Gold antifade regent; Invitrogen, Thermo Fisher Scientific, MA, US) and imaged.

    Techniques: Quantitative RT-PCR, Western Blot, Expressing

    Development of retinal, intersegmental vasculature and expression of Vegf-a and EGFP in 24 h post-fertilization (hpf) Tg(fli1:EGFP)y 1 zebrafish embryos treated with IMD0354. a Detection of EGFP signal from Tg( fli1 :EGFP)y 1 transgenic zebrafish embryos retinal vasculature at 72 hpf treated with DMSO or IMD0354 (5, 10 ng/ml). Yellow arrows indicate the retinal vessels. White arrow indicates OA (optic artery). b Quantification of the number of retinal vessels at 72 hpf with DMSO or IMD0354 treatment (5 and 10 ng/ml); ( n = 7) One-way ANOVA test with Tukey multiple comparisons was used to determine statistical significance. c Detection of EGFP signal from Tg( fli1 :EGFP)y 1 transgenic zebrafish embryos intersegmental vasculature at 28 hpf treated with DMSO or IMD0354 (5 and 10 ng/ml). d Quantification of intersegmental vessel length ( n = 16) One-way ANOVA test with Tukey multiple comparisons was used to determine statistical significance. e Immunofluorescent detection of Vegf-a (red) expression in zebrafish embryos at 24 hpf. f Western blot analysis of Vegf-a (monomeric and dimeric forms) and Egfp expression in the whole lysate of Tg(fli1:EGFP)y 1 transgenic zebrafish embryos at 24 hpf, incubated with DMSO and IMD0354 (10 and 5 ng/ml). β-Actin as the loading control. n.s. p > 0.05; *** p

    Journal: Angiogenesis

    Article Title: Selective IKK2 inhibitor IMD0354 disrupts NF-κB signaling to suppress corneal inflammation and angiogenesis

    doi: 10.1007/s10456-018-9594-9

    Figure Lengend Snippet: Development of retinal, intersegmental vasculature and expression of Vegf-a and EGFP in 24 h post-fertilization (hpf) Tg(fli1:EGFP)y 1 zebrafish embryos treated with IMD0354. a Detection of EGFP signal from Tg( fli1 :EGFP)y 1 transgenic zebrafish embryos retinal vasculature at 72 hpf treated with DMSO or IMD0354 (5, 10 ng/ml). Yellow arrows indicate the retinal vessels. White arrow indicates OA (optic artery). b Quantification of the number of retinal vessels at 72 hpf with DMSO or IMD0354 treatment (5 and 10 ng/ml); ( n = 7) One-way ANOVA test with Tukey multiple comparisons was used to determine statistical significance. c Detection of EGFP signal from Tg( fli1 :EGFP)y 1 transgenic zebrafish embryos intersegmental vasculature at 28 hpf treated with DMSO or IMD0354 (5 and 10 ng/ml). d Quantification of intersegmental vessel length ( n = 16) One-way ANOVA test with Tukey multiple comparisons was used to determine statistical significance. e Immunofluorescent detection of Vegf-a (red) expression in zebrafish embryos at 24 hpf. f Western blot analysis of Vegf-a (monomeric and dimeric forms) and Egfp expression in the whole lysate of Tg(fli1:EGFP)y 1 transgenic zebrafish embryos at 24 hpf, incubated with DMSO and IMD0354 (10 and 5 ng/ml). β-Actin as the loading control. n.s. p > 0.05; *** p

    Article Snippet: Samples were incubated with VEGF-a (GTX21316, 1:100; GeneTex, Simpson, PA, US) antibody, then visualized (DyLight 519, 1:500; Thermo Fisher Scientific, MA, US), mounted (ProLong Gold antifade regent; Invitrogen, Thermo Fisher Scientific, MA, US) and imaged.

    Techniques: Expressing, Transgenic Assay, Western Blot, Incubation

    Histology and immunofluorescence in rat corneal sections. a Hematoxylin and Eosin (H E), b VEGF-A (green); c CCL2 (green); d TNF-α (green); e CXCL5 (green); f CD45 (green) and g HIF-1α (green) staining in rat cornea tissue. Nuclear counterstaining by DAPI (blue) in fluorescent images

    Journal: Angiogenesis

    Article Title: Selective IKK2 inhibitor IMD0354 disrupts NF-κB signaling to suppress corneal inflammation and angiogenesis

    doi: 10.1007/s10456-018-9594-9

    Figure Lengend Snippet: Histology and immunofluorescence in rat corneal sections. a Hematoxylin and Eosin (H E), b VEGF-A (green); c CCL2 (green); d TNF-α (green); e CXCL5 (green); f CD45 (green) and g HIF-1α (green) staining in rat cornea tissue. Nuclear counterstaining by DAPI (blue) in fluorescent images

    Article Snippet: Samples were incubated with VEGF-a (GTX21316, 1:100; GeneTex, Simpson, PA, US) antibody, then visualized (DyLight 519, 1:500; Thermo Fisher Scientific, MA, US), mounted (ProLong Gold antifade regent; Invitrogen, Thermo Fisher Scientific, MA, US) and imaged.

    Techniques: Immunofluorescence, Staining

    Effect of IKK2 inhibition on HUVEC, microvessel outgrowth from aortic rings, and VEGF-A expression. a Dose-dependent (10, 5 and 2.5 ng/ml) inhibitory effect of IMD0354 on HUVEC migration relative to control (DMSO 1 µl/ml). Living HUVEC were visualized with Calcein-AM (green). b Quantification of HUVEC migration distance ( n = 8). One-way ANOVA test with Tukey multiple comparison was used to determine statistical significance. c HUVEC has grown on Geltrex to evaluate tube formation in the presence of IMD0354 compared to drug-free vehicle (DMSO). Vital HUVEC are stained with calcein-AM (green), and dead HUVEC are displayed in red (propidium iodide). Quantitative analysis of a number of junctions ( d ), and tubules ( e ) formed by HUVEC, and total tubule length ( f ) ( n = 8). g Quantitative analysis of cell death induced by IMD0354 treatment ( n = 8). h Effect of IMD0354 on cell proliferation in the aortic ring assay. Student t test was used to determine statistical significance. i Western blot analysis of VEGF-A expression in HUVEC treated with IMD0354 (10, 5, 2.5 ng/ml), with β-actin as a loading control. j Western blot analysis of HIF-1α expression in HUVEC treated with IMD0354 (10, 5, 2.5 ng/ml), with β-actin as a loading control. k Immunofluorescent detection of VEGF-A (green) in HUVEC treated with IMD0354 (10, 5, 2.5 ng/ml). Cell nuclei visualized with DAPI staining (blue). n.s. p > 0.05; * p

    Journal: Angiogenesis

    Article Title: Selective IKK2 inhibitor IMD0354 disrupts NF-κB signaling to suppress corneal inflammation and angiogenesis

    doi: 10.1007/s10456-018-9594-9

    Figure Lengend Snippet: Effect of IKK2 inhibition on HUVEC, microvessel outgrowth from aortic rings, and VEGF-A expression. a Dose-dependent (10, 5 and 2.5 ng/ml) inhibitory effect of IMD0354 on HUVEC migration relative to control (DMSO 1 µl/ml). Living HUVEC were visualized with Calcein-AM (green). b Quantification of HUVEC migration distance ( n = 8). One-way ANOVA test with Tukey multiple comparison was used to determine statistical significance. c HUVEC has grown on Geltrex to evaluate tube formation in the presence of IMD0354 compared to drug-free vehicle (DMSO). Vital HUVEC are stained with calcein-AM (green), and dead HUVEC are displayed in red (propidium iodide). Quantitative analysis of a number of junctions ( d ), and tubules ( e ) formed by HUVEC, and total tubule length ( f ) ( n = 8). g Quantitative analysis of cell death induced by IMD0354 treatment ( n = 8). h Effect of IMD0354 on cell proliferation in the aortic ring assay. Student t test was used to determine statistical significance. i Western blot analysis of VEGF-A expression in HUVEC treated with IMD0354 (10, 5, 2.5 ng/ml), with β-actin as a loading control. j Western blot analysis of HIF-1α expression in HUVEC treated with IMD0354 (10, 5, 2.5 ng/ml), with β-actin as a loading control. k Immunofluorescent detection of VEGF-A (green) in HUVEC treated with IMD0354 (10, 5, 2.5 ng/ml). Cell nuclei visualized with DAPI staining (blue). n.s. p > 0.05; * p

    Article Snippet: Samples were incubated with VEGF-a (GTX21316, 1:100; GeneTex, Simpson, PA, US) antibody, then visualized (DyLight 519, 1:500; Thermo Fisher Scientific, MA, US), mounted (ProLong Gold antifade regent; Invitrogen, Thermo Fisher Scientific, MA, US) and imaged.

    Techniques: Inhibition, Expressing, Migration, Staining, Aortic Ring Assay, Western Blot

    The NOR-1 protector prevents the down-regulation of VCAM-1 promoted by miR-17 and -20. HUVEC were transfected with precursors of either miR-17 or miR-20a (P17 or P20) or a scramble sequence (Scr) in the presence or in the absence of a miScript target protector that blocks the NOR-1 3’-UTR seed sequence recognized by the miR-17 and -20a (NOR1-Prot). Then cells were stimulated with VEGF (100 ng/ml, 2 h). VCAM-1 expression was assessed by real-time PCR (A) and Western-blot (B) . Results are expressed as mean ± SD from at least n = 4. ( p

    Journal: PLoS ONE

    Article Title: miR-17 and -20a Target the Neuron-Derived Orphan Receptor-1 (NOR-1) in Vascular Endothelial Cells

    doi: 10.1371/journal.pone.0141932

    Figure Lengend Snippet: The NOR-1 protector prevents the down-regulation of VCAM-1 promoted by miR-17 and -20. HUVEC were transfected with precursors of either miR-17 or miR-20a (P17 or P20) or a scramble sequence (Scr) in the presence or in the absence of a miScript target protector that blocks the NOR-1 3’-UTR seed sequence recognized by the miR-17 and -20a (NOR1-Prot). Then cells were stimulated with VEGF (100 ng/ml, 2 h). VCAM-1 expression was assessed by real-time PCR (A) and Western-blot (B) . Results are expressed as mean ± SD from at least n = 4. ( p

    Article Snippet: Finally, cells were stimulated with VEGF-A (100 ng/ml 2 h; R & D Systems).

    Techniques: Transfection, Sequencing, Expressing, Real-time Polymerase Chain Reaction, Western Blot

    A protector sequence against the NOR-1 3’-UTR seed region prevents the regulation by miR-17 and -20. HUVEC were transfected with precursors of either miR-17 or miR-20a (P17 or P20) or a scramble sequence (Scr) in the presence or in the absence of a miScript target protector that blocks the NOR-1 3’-UTR seed sequence recognized by the miR-17 and -20 (NOR1-Prot). Then cells were stimulated with VEGF (100 ng/ml, 2 h). NOR-1 expression was assessed by real-time PCR (A) and Western-blot (B) . Results are expressed as mean ± SD from at least n = 4. ( p

    Journal: PLoS ONE

    Article Title: miR-17 and -20a Target the Neuron-Derived Orphan Receptor-1 (NOR-1) in Vascular Endothelial Cells

    doi: 10.1371/journal.pone.0141932

    Figure Lengend Snippet: A protector sequence against the NOR-1 3’-UTR seed region prevents the regulation by miR-17 and -20. HUVEC were transfected with precursors of either miR-17 or miR-20a (P17 or P20) or a scramble sequence (Scr) in the presence or in the absence of a miScript target protector that blocks the NOR-1 3’-UTR seed sequence recognized by the miR-17 and -20 (NOR1-Prot). Then cells were stimulated with VEGF (100 ng/ml, 2 h). NOR-1 expression was assessed by real-time PCR (A) and Western-blot (B) . Results are expressed as mean ± SD from at least n = 4. ( p

    Article Snippet: Finally, cells were stimulated with VEGF-A (100 ng/ml 2 h; R & D Systems).

    Techniques: Sequencing, Transfection, Expressing, Real-time Polymerase Chain Reaction, Western Blot

    miR-17 and -20a regulate the expression of NOR-1. HUVEC were transfected with precursors of either miR-17 or miR-20a (P17 or P20) or a scramble sequence (Scr) in the presence or in the absence of their corresponding specific antagomirs (A17 or A20). Cells were stimulated with VEGF (100 ng/ml, 2 h). NOR-1 expression was assessed by real-time PCR (A) , Western-blot (B) and immunocytochemistry (C) . Results are expressed as mean ± SD from at least n = 4. ( p

    Journal: PLoS ONE

    Article Title: miR-17 and -20a Target the Neuron-Derived Orphan Receptor-1 (NOR-1) in Vascular Endothelial Cells

    doi: 10.1371/journal.pone.0141932

    Figure Lengend Snippet: miR-17 and -20a regulate the expression of NOR-1. HUVEC were transfected with precursors of either miR-17 or miR-20a (P17 or P20) or a scramble sequence (Scr) in the presence or in the absence of their corresponding specific antagomirs (A17 or A20). Cells were stimulated with VEGF (100 ng/ml, 2 h). NOR-1 expression was assessed by real-time PCR (A) , Western-blot (B) and immunocytochemistry (C) . Results are expressed as mean ± SD from at least n = 4. ( p

    Article Snippet: Finally, cells were stimulated with VEGF-A (100 ng/ml 2 h; R & D Systems).

    Techniques: Expressing, Transfection, Sequencing, Real-time Polymerase Chain Reaction, Western Blot, Immunocytochemistry

    miR-329 is downregulated by angiogenic factors resulting in increased CD146 expression. (A and B) HUVECs were cultured in the presence of VEGF (50 ng/ml) or TNF-α (50 ng/ml) or coincubated with NF-κB pathway inhibitor BAY11-7082 (200 ng/ml)

    Journal: Molecular and Cellular Biology

    Article Title: MicroRNA 329 Suppresses Angiogenesis by Targeting CD146

    doi: 10.1128/MCB.00343-13

    Figure Lengend Snippet: miR-329 is downregulated by angiogenic factors resulting in increased CD146 expression. (A and B) HUVECs were cultured in the presence of VEGF (50 ng/ml) or TNF-α (50 ng/ml) or coincubated with NF-κB pathway inhibitor BAY11-7082 (200 ng/ml)

    Article Snippet: Serum-starved HUVECs (12 h) were incubated with NF-κB inhibitor BAY11-7082 (200 ng/ml) or transfected with different small RNAs or vectors before induction with VEGF (Upstate Biotechnology) or TNF-α (Peprotech) (50 ng/ml each).

    Techniques: Expressing, Cell Culture

    Effect of DHA-supplemented diet on long-term diabetes-induced degenerative changes in rat retina. Retinas isolated 9 months after induction of diabetes were analyzed by quantitative PCR and immunobloting for inflammatory/angiogenic molecule expression. Quantitative PCR analysis of ASM, ICAM-1, VEGF, and IL-1β ( A ) of retinas isolated from rats subjected to a standard diet (control, white bar ; diabetic, black bar ) or a DHA-enriched diet (diabetic, striped bar) is shown. The results are means ± SE from one set of animals, with five to eight animals in each group. * P

    Journal: Diabetes

    Article Title: The Unconventional Role of Acid Sphingomyelinase in Regulation of Retinal Microangiopathy in Diabetic Human and Animal Models

    doi: 10.2337/db10-0550

    Figure Lengend Snippet: Effect of DHA-supplemented diet on long-term diabetes-induced degenerative changes in rat retina. Retinas isolated 9 months after induction of diabetes were analyzed by quantitative PCR and immunobloting for inflammatory/angiogenic molecule expression. Quantitative PCR analysis of ASM, ICAM-1, VEGF, and IL-1β ( A ) of retinas isolated from rats subjected to a standard diet (control, white bar ; diabetic, black bar ) or a DHA-enriched diet (diabetic, striped bar) is shown. The results are means ± SE from one set of animals, with five to eight animals in each group. * P

    Article Snippet: ASM antibody (Kolesnick laboratory), VEGF antibody (Novus Biologicals, Littleton, CO), caveolin-1 antibody (BD Bioscience, San Jose, CA), lysosomal associated membrane protein 1 (LAMP1) antibody (BD Bioscience), and ICAM-1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA) were used.

    Techniques: Isolation, Real-time Polymerase Chain Reaction, Western Blot, Expressing

    Effect of DHA-supplemented diet on short-term diabetes-induced changes in rat retina. Retinas isolated 1 month after induction of diabetes were analyzed by quantitative PCR and Western blotting for inflammatory/angiogenic molecule expression levels. Quantitative PCR analysis of ASM, ICAM-1, VEGF, and IL-1β ( A ) from retinas isolated from rats on standard diet (control, white bar ; diabetic, black bar) and a DHA-enriched diet (diabetic, striped bar) is shown. The results are means ± SE from three independent sets of animals, with five to eight animals in each group. * P

    Journal: Diabetes

    Article Title: The Unconventional Role of Acid Sphingomyelinase in Regulation of Retinal Microangiopathy in Diabetic Human and Animal Models

    doi: 10.2337/db10-0550

    Figure Lengend Snippet: Effect of DHA-supplemented diet on short-term diabetes-induced changes in rat retina. Retinas isolated 1 month after induction of diabetes were analyzed by quantitative PCR and Western blotting for inflammatory/angiogenic molecule expression levels. Quantitative PCR analysis of ASM, ICAM-1, VEGF, and IL-1β ( A ) from retinas isolated from rats on standard diet (control, white bar ; diabetic, black bar) and a DHA-enriched diet (diabetic, striped bar) is shown. The results are means ± SE from three independent sets of animals, with five to eight animals in each group. * P

    Article Snippet: ASM antibody (Kolesnick laboratory), VEGF antibody (Novus Biologicals, Littleton, CO), caveolin-1 antibody (BD Bioscience, San Jose, CA), lysosomal associated membrane protein 1 (LAMP1) antibody (BD Bioscience), and ICAM-1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA) were used.

    Techniques: Isolation, Real-time Polymerase Chain Reaction, Western Blot, Expressing

    Retinal gene expression and capillary degeneration in mice undergoing retinal I/R injury. Quantitative PCR analysis of ASM, ICAM-1, VCAM-1, IL-1β, TNF-α, and VEGF ( A ) in retinas isolated from ASM +/+ and ASM −/− mice 2 days after retinal I/R injury is shown. The data are presented as means ± SE from three independent sets of mice performed in triplicate with a total of five mice per group. B : Acellular capillary occurrence (black arrows) in the retina isolated from ASM +/+ and ASM −/− mice 7 days after retinal I/R injury. Quantification of the number of acellular capillaries from five independent sets of mice, with a total of seven mice per ASM +/+ group and six mice per ASM −/− group, is shown. At least eight fields of retina were counted in duplicates by two independent investigators. * P

    Journal: Diabetes

    Article Title: The Unconventional Role of Acid Sphingomyelinase in Regulation of Retinal Microangiopathy in Diabetic Human and Animal Models

    doi: 10.2337/db10-0550

    Figure Lengend Snippet: Retinal gene expression and capillary degeneration in mice undergoing retinal I/R injury. Quantitative PCR analysis of ASM, ICAM-1, VCAM-1, IL-1β, TNF-α, and VEGF ( A ) in retinas isolated from ASM +/+ and ASM −/− mice 2 days after retinal I/R injury is shown. The data are presented as means ± SE from three independent sets of mice performed in triplicate with a total of five mice per group. B : Acellular capillary occurrence (black arrows) in the retina isolated from ASM +/+ and ASM −/− mice 7 days after retinal I/R injury. Quantification of the number of acellular capillaries from five independent sets of mice, with a total of seven mice per ASM +/+ group and six mice per ASM −/− group, is shown. At least eight fields of retina were counted in duplicates by two independent investigators. * P

    Article Snippet: ASM antibody (Kolesnick laboratory), VEGF antibody (Novus Biologicals, Littleton, CO), caveolin-1 antibody (BD Bioscience, San Jose, CA), lysosomal associated membrane protein 1 (LAMP1) antibody (BD Bioscience), and ICAM-1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA) were used.

    Techniques: Expressing, Mouse Assay, Real-time Polymerase Chain Reaction, Isolation

    GNF351-mediated repression of growth factor expression is AHR-dependent in primary FLS. Repression of AHR expression was performed using AHR-specific siRNA in primary FLS from healthy donors (A) or from patients with RA (B). A total of 2 × 10 6 cells were used per gene knockdown sample. The repression of AHR protein expression was confirmed using Western blot analysis after 48 hours of siRNA transfection. Upon confirmation, primary N-FLS transfected with siRNA for 48 hours were pretreated with 500 nM GNF351 for 1 hour, followed by 10 ng/ml IL1B challenge for 4 hours. The levels of EREG , AREG , VEGF-A , and FGF-2 mRNA were determined by quantitative RT-PCR analysis. Data points and bars represent mean ± S.E. of three independent determinations. Data were analyzed using one-way ANOVA followed by Tukey’s multiple-comparisons test. * P

    Journal: The Journal of Pharmacology and Experimental Therapeutics

    Article Title:

    doi: 10.1124/jpet.113.209726

    Figure Lengend Snippet: GNF351-mediated repression of growth factor expression is AHR-dependent in primary FLS. Repression of AHR expression was performed using AHR-specific siRNA in primary FLS from healthy donors (A) or from patients with RA (B). A total of 2 × 10 6 cells were used per gene knockdown sample. The repression of AHR protein expression was confirmed using Western blot analysis after 48 hours of siRNA transfection. Upon confirmation, primary N-FLS transfected with siRNA for 48 hours were pretreated with 500 nM GNF351 for 1 hour, followed by 10 ng/ml IL1B challenge for 4 hours. The levels of EREG , AREG , VEGF-A , and FGF-2 mRNA were determined by quantitative RT-PCR analysis. Data points and bars represent mean ± S.E. of three independent determinations. Data were analyzed using one-way ANOVA followed by Tukey’s multiple-comparisons test. * P

    Article Snippet: To determine the levels of growth factor secretion, VEGF-A (Abnova, Taipei, Taiwan) and EREG (Uscn Life Science, Wuhan, China) ELISAs were performed per manufacturers’ instructions.

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

    GNF351 inhibits IL1B-induced expression of growth factors from primary FLS. Primary N-FLS (A) or RA-FLS (B) were treated with 500 nM GNF351 for 1 hour, followed by 4-hour stimulation with 10 ng/ml IL1B. The levels of EREG , AREG , VEGF-A , and FGF-2 mRNA were determined by quantitative RT-PCR analysis. Data points and bars represent mean ± S.E. of three independent determinations. Data were analyzed using one-way ANOVA followed by Tukey’s multiple-comparisons test. * P

    Journal: The Journal of Pharmacology and Experimental Therapeutics

    Article Title:

    doi: 10.1124/jpet.113.209726

    Figure Lengend Snippet: GNF351 inhibits IL1B-induced expression of growth factors from primary FLS. Primary N-FLS (A) or RA-FLS (B) were treated with 500 nM GNF351 for 1 hour, followed by 4-hour stimulation with 10 ng/ml IL1B. The levels of EREG , AREG , VEGF-A , and FGF-2 mRNA were determined by quantitative RT-PCR analysis. Data points and bars represent mean ± S.E. of three independent determinations. Data were analyzed using one-way ANOVA followed by Tukey’s multiple-comparisons test. * P

    Article Snippet: To determine the levels of growth factor secretion, VEGF-A (Abnova, Taipei, Taiwan) and EREG (Uscn Life Science, Wuhan, China) ELISAs were performed per manufacturers’ instructions.

    Techniques: Expressing, Quantitative RT-PCR

    AHR occupancy at the growth factor promoters drives their expression. (A) COS-1 cells were transfected with VEGF-A-HSV-TK-Luc, EREG-HSV-TK-Luc, or AREG-HSV-TK-Luc and pcDNA3-hAHR for 24 hours. Cells were treated with vehicle or 10 nM TCDD for 4 hours, followed by luciferase activity measurement. (B) For the VEGF-A and EREG ELISAs, primary RA-FLS cells were plated at 2 × 10 6 cells per ml. The cells were treated for 1 hour with 500 nM GNF351, followed by cytokine challenge with 10 ng/ml IL1B. Cells were retreated with GNF351 every 12 hours after IL1B treatment of 48 hours. Data points and bars represent mean ± S.E. of three independent determinations. Data were analyzed using one-way ANOVA followed by Tukey’s multiple-comparisons test. * P

    Journal: The Journal of Pharmacology and Experimental Therapeutics

    Article Title:

    doi: 10.1124/jpet.113.209726

    Figure Lengend Snippet: AHR occupancy at the growth factor promoters drives their expression. (A) COS-1 cells were transfected with VEGF-A-HSV-TK-Luc, EREG-HSV-TK-Luc, or AREG-HSV-TK-Luc and pcDNA3-hAHR for 24 hours. Cells were treated with vehicle or 10 nM TCDD for 4 hours, followed by luciferase activity measurement. (B) For the VEGF-A and EREG ELISAs, primary RA-FLS cells were plated at 2 × 10 6 cells per ml. The cells were treated for 1 hour with 500 nM GNF351, followed by cytokine challenge with 10 ng/ml IL1B. Cells were retreated with GNF351 every 12 hours after IL1B treatment of 48 hours. Data points and bars represent mean ± S.E. of three independent determinations. Data were analyzed using one-way ANOVA followed by Tukey’s multiple-comparisons test. * P

    Article Snippet: To determine the levels of growth factor secretion, VEGF-A (Abnova, Taipei, Taiwan) and EREG (Uscn Life Science, Wuhan, China) ELISAs were performed per manufacturers’ instructions.

    Techniques: Expressing, Transfection, Luciferase, Activity Assay

    Immunohistochemical staining of high and low expression of COX-2 ( A , D ), EGFR ( B , E ), and VEGF ( C , F ).

    Journal: Journal of Korean Medical Science

    Article Title: VEGF as a Predictor for Response to Definitive Chemoradiotherapy and COX-2 as a Prognosticator for Survival in Esophageal Squamous Cell Carcinoma

    doi: 10.3346/jkms.2011.26.4.513

    Figure Lengend Snippet: Immunohistochemical staining of high and low expression of COX-2 ( A , D ), EGFR ( B , E ), and VEGF ( C , F ).

    Article Snippet: The slides were incubated overnight at 4℃ with the monoclonal antibodies, i.e., with the monoclonal antibody against EGFR (clone H11, diluted 1:200, Dako, Glostrup, Denmark), VEGF (clone VG1, diluted 1:50, Dako), and COX-2 (clone CX-294, diluted 1:100, Dako).

    Techniques: Immunohistochemistry, Staining, Expressing

    Overall survival according to the treatment response and the patterns of protein expression: ( A ) The Kaplan-Meier survival curve shows a significant difference between the patients with a complete response (CR) and those without a complete response. ( B , C ) the survival analyses of EGFR ( B ) and VEGF ( C ) did not show significant differences. ( D ) The patients with a higher expression of COX-2 show a significantly lower overall survival.

    Journal: Journal of Korean Medical Science

    Article Title: VEGF as a Predictor for Response to Definitive Chemoradiotherapy and COX-2 as a Prognosticator for Survival in Esophageal Squamous Cell Carcinoma

    doi: 10.3346/jkms.2011.26.4.513

    Figure Lengend Snippet: Overall survival according to the treatment response and the patterns of protein expression: ( A ) The Kaplan-Meier survival curve shows a significant difference between the patients with a complete response (CR) and those without a complete response. ( B , C ) the survival analyses of EGFR ( B ) and VEGF ( C ) did not show significant differences. ( D ) The patients with a higher expression of COX-2 show a significantly lower overall survival.

    Article Snippet: The slides were incubated overnight at 4℃ with the monoclonal antibodies, i.e., with the monoclonal antibody against EGFR (clone H11, diluted 1:200, Dako, Glostrup, Denmark), VEGF (clone VG1, diluted 1:50, Dako), and COX-2 (clone CX-294, diluted 1:100, Dako).

    Techniques: Expressing