Federal government websites often end in .gov or .mil. Erg prevents NF-B p65 from binding to the ICAM-1 promoter, suggesting a direct mechanism of interference. designed and carried out invitro experiments, analyzed, interpreted, and conceptualized results, and wrote the manuscript. Immunohistochemistry analysis revealed endothelial cells stained positive for ERG, D2-40 (podoplanin) and negative for WT1, calretinin. Endothelial cytoplasm is inconspicuous in routine light microscopy. Identification et impacts des anomalies gntiques dans la gense, l'volution clinique et le traitement des gliomes (C) -catenin (-cat; green) and VE-cadherin (VEC; red) staining of FITC-conjugated siCtrl and siERG (FITC; purple) treated HUVEC (n= 3). (A) Western blot of -catenin expression in control and ERG-deficient cells treated in presence or absence of MG132 (n= 4). 2012; 124: 763775. (F) Quantification of pericyte coverage, pixel intensity (n= 8). Analysis of the promoter sequences of other genes highlighted in the GSEA identified putative Erg binding sites within a distance equivalent to 12 nucleosomes away from an NF-B site.3 The involvement of distant sites in transcriptional repression has been well described, and may involve recruitment of co-repressors and/or modification of chromatin structure (41). ( C) Endothelial cells ERG positive. The Wnt/beta-catenin pathway modulates vascular remodeling and specification by upregulating Dll4/Notch signaling. ETS-related gene (ERG) is a transcription factor that has been linked to angiogenesis. However, the EBS mutant constructs 118 and 181, and the combined 118/181 mutant, showed no significant increase in promoter activity after Erg Genebloc treatment compared with control Genebloc, indicating involvement of these sites in Erg-mediated repression of ICAM-1 promoter activity. June 13, Mice were administered Tamoxifen (50g per mouse; Sigma) by intraperitoneal injection (IP) at postnatal (P) day 1, P2 and P3. The chromatin-remodeling enzyme BRG1 modulates vascular Wnt signaling at two levels. Inactivation of AR/TMPRSS2-ERG/Wnt signaling networks attenuates the aggressive behavior of prostate cancer cells. Federal government websites often end in .gov or .mil. Deletion of either VEGF-A or VEGFR2 in endothelial cells regulates levels of Dll4 in tip cells and, in turn, Notch inhibition provides a feedback loop that reinforces expression of VEGF-A and C-X-C chemokine receptor type 4 (CXCR4), which stimulate endothelial sprouting and proliferation in the expanding vascular plexus [13 ]. and transmitted securely. Transcription factor Erg regulates angiogenesis and endothelial apoptosis through VE-cadherin. Shifted protein-oligonucleotide complexes are indicated by an arrow and super-shifted complexes are indicated by arrowhead. Careers. To further define the role of ERG in regulating EC function, we evaluated the effect of ERG knockdown on EC lumen formation in 3D collagen matrices. Angiogenesis of glioma: evaluation of ultrastructural characteristics of microvessels and tubular bodies (Weibel-Palade) in endothelial cells and immunohistochemical findings with VEGF and p53 protein. The site is secure. Chromatin was immunoprecipitated with 2 g antibody to Erg (sc-353, Santa Cruz Biotechnology, Inc), NF-B p65 (ab7970, AbCam, Cambridge, United Kingdom), or negative control rabbit IgG (PP64, Chemicon, Millipore). In addition, we also used GSEA to compare the Erg-regulated genes with the data from a recent ChIP seq analysis, which identified regulatory sequences bound by NF-B p65 in LPS-treated macrophages (19). Location of quantitative PCR amplicons covering R1, R2, R3, R4, and R5 are indicated by black lines below. Addition of an anti-Erg antibody resulted in the appearance of a supershift (Fig. Please enable it to take advantage of the complete set of features! 2015 The Authors. Wnt/beta-catenin signaling controls development of the blood-brain barrier. 5B). Dustri-Verlag Dr. Karl Feistle GmbH & Co. KG. Therapeutic angiogenesis for cardiovascular disease: biological context, challenges, prospects. Cell 32, 8296 (2015). In endothelial cells (EC), the transcription factor NF-B is important in the switch from quiescence to activation, and is tightly controlled to avoid excessive inflammation and organ damage. CD31 demonstrated variable and sometimes weak immunoreactivity for endothelial cells. Nuclear lysates from a confluent monolayer of HUVEC were extracted using the Nuclear extract kit (Active Motif) following the manufacturer's instructions. Dynamic regulation of canonical TGFbeta signalling by endothelial transcription factor ERG protects from liver fibrogenesis. To confirm that ERG controls angiogenesis and vascular development in a Wnt/-catenin-dependent manner invivo, we carried out a rescue experiment by pharmacological stabilization of Wnt/-catenin signaling (. E ndothelial cells (EC) have many functions and play a central role in the control of coagulation, thrombolysis, vascular tone, permeability, inflammation, tissue repair, and angiogenesis. By using human umbilical vein endothelial cells (HUVECs), we showed that Erg overexpression by adenovirus (AdErg) repressed basal and TNF--induced expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule (VCAM), and interleukin 8 (IL-8). eCollection 2022. MicroRNA-147a Targets SLC40A1 to Induce Ferroptosis in Human Glioblastoma. E26 transformation-specific (ETS)-related gene (ERG) belongs to the ETS transcriptional factor family and is required for endothelial cells (ECs) homeostasis and cardiac . Published by Elsevier Inc. We use cookies to help provide and enhance our service and tailor content. Nat Rev Cardiol. Bookshelf A. Endothelial Cells. In EC, Erg expression is down-regulated by inflammatory stimuli (9, 13); this is in contrast with ETS factors including Ets-1, Ets-2, and ESE-1, whose expression is increased after treatment with agents such as IL-1, TNF-, angiotensin II, or thrombin (9, 17, 5459). This article describes the possibility of direct reprogramming of non-vascular cells into endothelial cells using ETV2, ERG and FLI1 transcription factors. In this study we investigate the mechanisms used by Erg to repress inflammatory gene expression in quiescent EC, focusing on ICAM-1 as a model gene. 2014, Received: Dynamic Phosphorylation of CENP-A at Ser68 Orchestrates Its Cell-Cycle-Dependent Deposition at Centromeres, Reverse Genetic Screening Reveals Poor Correlation between Morpholino-Induced and Mutant Phenotypes in Zebrafish, Endothelial ERG Is Required for Vascular Development, Angiogenesis, and Tumor Growth, ERG Is Required for Vascular Development, Physiological Postnatal Angiogenesis, and Pathological Tumor Angiogenesis, ERG Controls Vascular Stability and Pericyte Coverage, ERG Controls -Catenin Stability and Signaling through VE-Cadherin- and Wnt-Dependent Mechanisms, Endothelial Canonical Wnt Signaling and -Catenin Stability Are Regulated by ERG, ERG Controls -Catenin Stability through VE-Cadherin- and Wnt-Dependent Mechanisms, Expression of the Wnt Receptor Frizzled-4 Is Regulated byERG, Baylor College of Medicine Human Genome Sequencing Center, Washington University Genome Sequencing Center, Childrens Hospital Oakland Research Institute, ERG Controls Angiogenesis through Wnt Signaling, ERG Regulates Angiogenesis through Wnt/-Catenin Signaling, ERG Overexpression Stabilizes VEGF-Induced Blood Vessels and Promotes Angiogenesis InVivo, Isolation of Mouse Lung Endothelial Cells, Plasmid Transfections and Reporter Assays, eyJraWQiOiI4ZjUxYWNhY2IzYjhiNjNlNzFlYmIzYWFmYTU5NmZmYyIsImFsZyI6IlJTMjU2In0.eyJzdWIiOiI0MTE4MjEwN2VhMzAwMjI5Y2M1YTE0ZWY1ZDllNjA4OCIsImtpZCI6IjhmNTFhY2FjYjNiOGI2M2U3MWViYjNhYWZhNTk2ZmZjIiwiZXhwIjoxNjcwODIyNzE3fQ.n2t_EEIadOWDuBvYBM3qxbgnXUkqeUfJTf3R687qKZj_pnMqh3eIgs7tPeGSFW6wtKoPQsvfeQ2e5TaMM76aisnz580wH5QVG1dLFDmHCQpgnnDOct2_VHnSBX1CGX83q7xhXMGqUOUE-HNIfQxZ53P5GwfcnHh1lLJU74cT2FkyUg4J_mvbMyU9e2FhBmwbjsBfMQeAEIM0gKLuvF4lAhAd_IXnZlTR5ly4rlairqiIeoKF1qr6Q53jhqOvogcfAuJtwEenI0cy2VsXzNimB5zlXFGUZGkjHhhrf0_KiHdfZfw9C0ffYD0fg93UcxK4QXJzlX7g01j7zDxB91xtIw, eyJraWQiOiI4ZjUxYWNhY2IzYjhiNjNlNzFlYmIzYWFmYTU5NmZmYyIsImFsZyI6IlJTMjU2In0.eyJzdWIiOiI4ZjMzNWYwNjIyYjJmYmIzZmM0Y2M4ZGYwYzY5MzJmNCIsImtpZCI6IjhmNTFhY2FjYjNiOGI2M2U3MWViYjNhYWZhNTk2ZmZjIiwiZXhwIjoxNjcwODIyNzE3fQ.idar2V0x9cfoVI0p0kPMyjqj1i5F1H25HkaKd9fNUWIKAtgAtdUoD6q69na4nkTzLP-H2cJzVfx2bpwJOyx-S1IpEqKf0-YyNA0Nea9nSSHrbqGDpCi6GHWiUSM-V27KBzit9SsbsWal6a1aQMSbDQL1S_Y9POeP40CfrC_s4Pe6PAz-Gagozm5_MCh4FenMF6DVpiL0Ts3Vvz85NVmt0how8309XLFsJL_IrMuonIQjU6L71zYGkwTTdoU4D1JNyfLkSAxnoKAAPNkfZM61-fW0sLo7omTLvNNhAJQS7kv7ofGuPMeCa6yNbR7Q6Qa4ENhb7HQuQTRdq5xZKyhFew, https://doi.org/10.1016/j.devcel.2014.11.016, The Endothelial Transcription Factor ERG Promotes Vascular Stability and Growth through Wnt/-Catenin Signaling, View Large Projects in the group focus on three main areas: - Transcriptional and epigenetic control of endothelial homeostasis by the ETS transcription factor ERG - von Willebrand Factor regulation of angiogenesis and angiodysplasia The following day, cells were transduced with 100 multiplicity of infection of IB Super Repressor Adenovirus (AdIBSR) (14) or AdLacZ in serum-free M199 medium for 2 h before replacing with complete M199 medium. These cells capture visual information and ultimately respond to light . We show that in quiescent EC Erg prevents NF-B p65 binding to DNA, suggesting that Erg may compete with p65 for DNA binding. Primers for site-directed mutagenesis are listed in supplemental Table S1. (A) Representative whole mount images of E10.5, (B) Endomucin staining of blood vessels in E10.5, (C) Isolectin B4 staining of postnatal day 6 retinas from. 2021 Jan 1;14(1):116-125. eCollection 2021. EIF5 EIF6 ELF2 ELK1 ELK3 EMC8 EME1 EMG1 ENKD1 ENO3 ENTR1 ENY2 EOGT EP400P1 EPB41 EPB41L1 EPB41L2 EPB41L4A EPB41L5 EPHA2 EPHB4 EPS8L2 ERC1 ERCC1 ERG ERH ERMN ERVFRD-1 ESRRG ETFB ETHE1 ETNPPL ETS1 ETV3 ETV4 ETV5 EVI2A EXOC3 EXOC6 EXOSC1 EXOSC3 EXOSC7 F12 FAAH2 FAAP100 . Retinas were collected at P6 and processed as described (. S3). ICAM-1 promoter activity is repressed after Erg overexpression in HUVEC (12); therefore we investigated whether this repression was lost after mutation of EBS in the ICAM-1 promoter. official website and that any information you provide is encrypted The role of Erg as a repressor of inflammation is in contrast to that of other ETS factors, which have previously been shown to act synergistically with NF-B in promoting inflammatory gene expression. A role for the beta-catenin/T-cell factor signaling cascade in vascular remodeling. The above data suggest that Erg is repressing ICAM-1 through a mechanism that involves EBS 118 and EBS 181. Temporal control and selective gene activation, Sun S. C., Ganchi P. A., Ballard D. W., Greene W. C. (1993), NF-B controls expression of inhibitor IB. Identification of the DNA binding site involved in Erg-mediated repression of ICAM-1. 8600 Rockville Pike Med Mol Morphol. 2001;161:III-XIII, 1-151. doi: 10.1007/978-3-642-56553-3. Furthermore, PGA-PTX-E-[c(RGDfK)2] blocked endothelial cells' migration towards vascular endothelial growth factor, and capillary-like tube formation, and inhibited endothelial cells' attachment to fibrinogen in the 4T1 breast tumor model. NF-B activity is tightly regulated through feedback repressive mechanisms (3, 4). After transduction with AdIBSR, the increase in ICAM-1 mRNA expression following Erg inhibition was lost, compared with cells transduced with AdlacZ (Fig. The https:// ensures that you are connecting to the For normalization, tubulin was used as a cytoplasmic control and HDAC1 as a nuclear marker (n= 3). (2010), Genome-wide analysis of ETS family DNA-binding, Wilson N. K., Foster S. D., Wang X., Knezevic K., Schtte J., Kaimakis P., Chilarska P. M., Kinston S., Ouwehand W. H., Dzierzak E., Pimanda J. E., de Bruijn M. F., Gttgens B. Lack of pericytes leads to endothelial hyperplasia and abnormal vascular morphogenesis. Rafii, S., Butler, J. M. & Ding, B.-S. Angiocrine functions of organ-specific endothelial cells. Analysis of the IL-8 promoter showed that the EBS identified as the binding site of Erg is located within the functional NF-B binding site, similarly to what was observed for ICAM-1 and cIAP2 (supplemental Fig. There is growing evidence on the importance of transcription factor Erg in maintaining EC homeostasis. Numerous studies have shown that the new vasculature induced by VEGF invivo, to promote revascularization in ischemic diseases (. Other ETS combinatorial transcription factor motifs include the serum response elements that bind ternary complex ETS factors and the serum response factor (45), and ETS:AP1 binding motifs (46). This is likely to be the result of endothelial activation by proinflammatory stimuli, because Erg levels have been shown to decrease upon LPS or TNF- stimulation (9, 13). (D) Vascular density of isolectin B4 stained branches in the central plexus, scale bar, 50m; quantification (n= 6). Our previous studies have shown that ERG expression is highly enriched in endothelial cells (EC) both in vitro and in vivo. Briefly, biotinylated double-stranded oligonucleotides containing the ICAM-1 promoter sequence surrounding and including EBS 118 and 181 were incubated with 2 g of HUVEC nuclear lysate with or without excess unbiotinylated oligonucleotides. (E) (Top) Representative whole mount images of E10.5. 3A, lane 1, arrows), two of which were competed by excess unlabeled probe but not by a probe containing a mutation in the EBS 118 site, indicating specificity for this site. Our results show that ERG is a novel, reliable, and specific marker for endothelial cells within CNS tumors that can be used to better study the process of neovascularization. Regulation Of Endothelial Cell Migration; . Enhanced pathological angiogenesis in mice lacking beta3 integrin or beta3 and beta5 integrins. Nrarp coordinates endothelial Notch and Wnt signaling to control vessel density in angiogenesis. A, schematic diagram of ICAM-1 promoter mutant constructs. Results are expressed as fold-change compared with IgG normalized to input and negative control region. -, Hardee ME Zagzag D Mechanisms of glioma-associated neovascularization. (BD) (B) Representative images of EC sprouts on fibrin gel beads using siCtrl or siERG-treated HUVEC in the presence or absence of LiCl; (C) quantification of numbers of sprouts; and (D) tube length (n= 20). These data indicate that Erg binds to the ICAM-1 promoter in a region detected by the R4 primers, 188 to 103 bp upstream of the transcription start site. Cell sorting and proliferation assays performed after sustained ERG knockdown indicate that ERG drives proliferation and blocks the differentiation of prostate cells to both NE and luminal cell types. The identification of key mediators to regain control of EC homeostasis has great potential for the development of novel therapeutics. Our data highlights for the first time a mechanism of combinatorial repression involving Erg and NF-B, controlling basal repression of ICAM-1 and other proinflammatory endothelial genes. Kimble AL, Silva J, Omar OM, Murphy M, Hensel JA, Nicholas SE, Jellison ER, Reese B, Murphy PA. J Clin Pathol. 2022 Sep 16;23(18):10848. doi: 10.3390/ijms231810848. In this study we identify a novel mechanism that controls endothelial quiescence through inhibition of NF-B activity. The graphical outputs show enrichment (green curve) of genes up-regulated in TNF--treated pancreatic cancer cells (A), endothelial cells (B), or genes bound by NF-B p65 in macrophages after LPS stimulation (C), along a ranked list of genes up- or down-regulated by 48 h of Erg inhibition. 3C); however, NF-B p65 was enriched in R4 after treatment with TNF-, as expected (34) (Fig. Antibodies to Erg, p65, or IgG were incubated with nuclear extract-oligonucleotide complexes as indicated. Zhang, X., Hu, C., Yuan, Y.-P., Song, P., Kong, C.-Y., Wu, H.-M., Tang, Q.-Z. Optimized fibrin gel bead assay for the study of angiogenesis. (G and H) Panels show endomucin staining of blood vessels in B16F0 tumors and the quantification of the number of endomucin-positive vessels, (n= 6), scale bar, 50m. Alternatively, after 42 h following adenovirus transduction, cells were treated with 10 ng/ml of TNF- for 6 h. ICAM-1 mRNA levels were assessed by quantitative RT-PCR, normalized to GAPDH. pGL4 ICAM-1 1.3 was mutated in either single or double EBS or in the NF-B binding site. The enhancers of endothelial specific genes are synergistically activated by FOX and ETS transcription factors containing FOXO:ETS motifs (44). Transcription factor Erg regulates angiogenesis and endothelial apoptosis through VE-cadherin. Internet Explorer). Thus, using two separate inhibitors of NF-B, we have demonstrated that the up-regulation of ICAM-1 after Erg inhibition is mediated by the activity of NF-B. To update your cookie settings, please visit the. D, panel ii, ChIP was carried out on sheared chromatin from confluent resting HUVEC, using an anti-Erg or control IgG antibody. P30 CA016087/CA/NCI NIH HHS/United States, 1R21NS074055-01/NS/NINDS NIH HHS/United States, R01CA100426-01A1/CA/NCI NIH HHS/United States, Zagzag D Zhong H Scalzitti JM Laughner E Simons JW Semenza GL Expression of hypoxia-inducible factor 1alpha in brain tumors: association with angiogenesis, invasion, and progression. 2014, Received in revised form: Sci-Hub | Downregulation of ERG and FLI1 expression in endothelial cells triggers endothelial-to-mesenchymal transition. To expand this analysis to the entire Erg dataset and find patterns that could support the biological data, we used GSEA (3638), and compared the Erg dataset with other relevant gene sets, to identify common biological functions. Crucial among these are the adhesion molecule vascular endothelial (VE)-cadherin and its intracellular partner -catenin, an essential component of the canonical Wnt pathway (reviewed in. HUVEC were seeded onto 1% gelatin-coated plates and grown in EGM-2 medium (Lonza, Wokingham, United Kingdom). He was discharged from our hospital on postoperative day 6. The different expression levels and activities of ETS factors highlights their important role in regulating inflammation. and JavaScript. Zhou N, Ye Y, Wang X, Ma B, Wu J, Li L, Wang L, Wang DW, Zou Y. J Mol Med (Berl). 2022 Dec;58:102513. doi: 10.1016/j.redox.2022.102513. Immunoprecipitated DNA was analyzed by qPCR for primers covering EBS in cIAP2 (see supplemental Fig. The cyclin D1 gene is a target of the beta-catenin/LEF-1 pathway. Inhibition of Erg in resting HUVEC stimulates leukocyte recruitment (13). Don't show this again. Recently, we and others have shown that Erg represses endothelial expression of proinflammatory molecules ICAM-1 and IL-8 in quiescent cells, and that inhibition of Erg induces leukocyte adhesion to unstimulated human umbilical vein endothelial cells (HUVEC) (12, 13), suggesting an important role for Erg in maintaining EC homeostasis by repressing basal expression of proinflammatory genes. HHS Vulnerability Disclosure, Help Genome-wide analysis of the zebrafish ETS family identifies three genes required for hemangioblast differentiation or angiogenesis. NF-B induces ICAM-1 expression after Erg inhibition. (2003), Responses to the proinflammatory cytokines interleukin-1 and tumor necrosis factor in cells derived from rheumatoid synovium and other joint tissues involve nuclear factor B-mediated induction of the Ets transcription factor ESE-1, Creative Commons Attribution Non-Commercial License. 2015 Jan;68(1):44-50. doi: 10.1136/jclinpath-2014-202629. Moreover we showed that overexpression of Erg inhibited acute inflammation in mouse paws induced by TNF- (12). (1997), ETS1, NFB, and AP1 synergistically transactivate the human GM-CSF promoter, Gri G., Savio D., Trinchieri G., Ma X. Differentiated chondrocytes showing stem cell markers indicate the transit-amplifying nature of Studies have also demonstrated that the synovium these cells but their significance in cartilage regen-and infrapatellar fat pad contain mesenchymal eration is still unclear [Blanpain et al. 7): Fli-1, the ETS factor with the closest homology to Erg; Ets-2, a more distantly related ETS factor known to interact with Erg (20); and GABP, which can act as an activator or repressor of transcription (21, 22). Get time limited or full article access on ReadCube. In endothelial cells (EC), the transcription factor NF-B is important in the switch from quiescence to activation, and is tightly controlled to avoid excessive inflammation and organ damage. The transcription factor Erg is essential for definitive hematopoiesis and the function of adult hematopoietic stem cells. 2010 Jun 8;121(22):2407-18. doi: 10.1161/CIRCULATIONAHA.110.938217. Depending on the tissue and organ, endothelial cells can vary from each other. In resting endothelial cells (EC),2 NF-B is sequestered in the cytoplasm by proteins of the inhibitor of NF-B (IB) family. 2021 Feb 17;11(2):138. doi: 10.3390/jpm11020138. Cells were transfected with control pCMV6 or pCMV6-Fzd4 plasmids and transduced with VEC-GFP adenovirus (n= 3). In this study, we identify a transcriptional program regulated by ERG that controls vascular stability and growth through the Wnt/-catenin pathway, in both a physiological and pathological context. Because Erg inhibits ICAM-1 expression by repressing NF-B activity, we hypothesized that Erg may also inhibit the expression of other NF-B target genes in resting endothelium. Epub 2014 Oct 28. (1995), de Launoit Y., Audette M., Pelczar H., Plaza S., Baert J. L. (1998), The transcription of the intercellular adhesion molecule-1 is regulated by Ets transcription factors, Yockell-Lelivre J., Spriet C., Cantin P., Malenfant P., Heliot L., de Launoit Y., Audette M. (2009), Functional cooperation between Stat-1 and ets-1 to optimize, Maurer P., T'Sas F., Coutte L., Callens N., Brenner C., Van Lint C., de Launoit Y., Baert J. L. (2003), FEV acts as a transcriptional repressor through its DNA-binding ETS domain and alanine-rich domain, Wei G. H., Badis G., Berger M. F., Kivioja T., Palin K., Enge M., Bonke M., Jolma A., Varjosalo M., Gehrke A. R., Yan J., Talukder S., Turunen M., Taipale M., Stunnenberg H. G., Ukkonen E., Hughes T. R., Bulyk M. L., Taipale J. D and E, ChIP was carried out using an anti-Erg or control IgG antibody on sheared chromatin from confluent resting HUVEC (D) or HUVEC treated with Erg or control siRNA (E). 3B, lane 11 compared with lane 1). The following day, siRNA (10 nm) was mixed with AtuFect01 lipid (1 g/ml, Silence Therapeutics) at 5 times concentration in Opti-MEM (Invitrogen), then added to cells for 24 or 48 h. Erg overexpression was carried out using a V5-tagged Erg-3 adenovirus (AdErg), as described previously (12). Pisano C, Terriaca S, Scioli MG, Nardi P, Altieri C, Orlandi A, Ruvolo G, Balistreri CR. Epub 2022 Oct 22. (G) qPCR of downstream -catenin target gene expression in control and ERG-deficient HUVEC: Cyclin D1, Axin-2, and TCF-1 (n= 4). Document S1. Endothelins in cardiovascular biology and therapeutics. J Pers Med. More importantly, we proved that endothelial ERG overexpression notably prevented pressure overload-induced cardiac fibrosis. Inducible gene targeting in the neonatal vasculature and analysis of retinal angiogenesis in mice. S3) and negative control GAPDH promoter region. To confirm the specificity for Erg binding at this site, ChIP was carried out on chromatin from HUVEC treated with Erg siRNA. Specificity was measured by addition of saturating amounts of competing oligonucleotide (competitor) or competing oligonucleotide with a mutation in the EBS 118 (A) or 181 (B) (M-competitor). Little is known about the maintenance of endothelial cell fate in adults. ETS factors regulate specific target genes through combinatorial promoter motifs and interaction with other transcription factors. Combined genomic and antisense analysis reveals that the transcription factor Erg is implicated in endothelial cell differentiation. b: ERG exclusively, Figure 6.. Metastatic carcinoma. Xu P, Ge FH, Li WX, Xu Z, Wang XL, Shen JL, Xu AB, Hao RR. Bethesda, MD 20894, Web Policies The patient had an uneventful post-operative recovery. FZD4 as a mediator of ERG oncogene-induced WNT signaling and epithelial-to-mesenchymal transition in human prostate cancer cells. Sign up for the Nature Briefing newsletter what matters in science, free to your inbox daily. As expected, inhibition of Erg expression by siRNA decreased the amount of Erg binding to the ICAM-1 promoter (see Fig. [BMC Molecular and Cell Biology] Full Article Published In Prostate Cell News Analysis of oligonucleotide screening and ChIP sequencing (ChIP Seq) data has suggested a possible specific Erg consensus motif of (A/C)GGAA(G/A) (28) or AGGA(A/T)(G/A) (29). a: H & E demonstrates markedly diffuse microvascular proliferation. 3B, lanes 710); however, contrary to what was observed for the 118 EMSA, no supershift was detectable. Erg is a member of the ETS family of transcription factors, characterized by a conserved ETS DNA binding domain that binds to a core consensus motif of GGA(A/T) (6). We show that Erg binds to the promoters of a number of NF-B target genes, repressing their basal expression. RGC death occurs by apoptosis or necrosis. performed and supervised the tumor angiogenesis experiments, analyzed results, and contributed to scientific discussion. No enrichment for NF-B p65 was found in R4, which contains the NF-B binding sites, EBS 181 and 118, or in flanking R3 and R5 (Fig. Caught up in a Wnt storm: Wnt signaling in cancer. The LNCaP-androgen dependent cell line was cultured and passaged 60 times over 16 months. 5C), indicating a trend toward a correlation. Disclaimer, National Library of Medicine Crucially, we demonstrate that overexpression ofERG invivo enhances vascular endothelial growth factor (VEGF)-dependent angiogenesis and promotes stability of VEGF-induced new blood vessels. For Apelin expression assay, cells were cultured in the presence of both 4-hydroxytamoxifen and recombinant human VEGF. Epub 2019 Jun 10. Erg enrichment at R4, which includes EBS 118 and EBS 181, was greater than at other regions containing EBS (Fig. All graphical data are SEM, To investigate the role of ERG in physiological and pathological postnatal angiogenesis, floxed. sharing sensitive information, make sure youre on a federal ChIP was carried out on a confluent monolayer of quiescent or TNF--treated HUVEC. The supershift was specific, as it was competed off by an excess of unlabeled probe, but not by unlabeled probe containing a mutation of the EBS 118 (Fig. 1E). (I) TCF reporter (TOP) activity in control and ERG-deficient HUVEC treated with rWnt3a. ERG and nestin: useful markers of immature vessels and novel prognostic markers in renal cell carcinoma. Dhaun N, Webb DJ. Wnt/beta-catenin signaling induces proliferation, survival and interleukin-8 in human endothelial cells. Bethesda, MD 20894, Web Policies 3D Reconstruction of Neovessels inside Matrigel Plugs Supplemented with VEGF and Adenovirus Expressing ERG, Related to Figure6. HUVEC were seeded at 3 104 per well in EGM-2 medium (Lonza) on a gelatin-coated 24-well plate, and the following day either transduced with AdErg or AdLacZ (50 multiplicity of infection), or transfected with 100 nm Erg or control Genebloc and maintained in EGM-2 medium. is a recipient of a DOC-fFORTE fellowship of the Austrian Academy of Sciences at the London Research Institute. C and D, ChIP was carried out on sheared chromatin from confluent resting HUVEC TNF (10 ng/ml for 30 min) (C), or HUVEC treated with Erg or control siRNA (D) using an anti-NF-B p65 or control IgG antibody. The oligonucleotide containing EBS 118 formed a number of complexes with nuclear proteins from resting HUVEC (Fig. and transmitted securely. This indicates a correlation between NF-B pathway target genes in endothelial cells and genes up-regulated by Erg inhibition in HUVEC. 2004;101(11):391520. n = 6 (B), n = 5 (C), *, p < 0.05; **, p < 0.01; ***, p < 0.001. This modality has been successfully applied to destruct ERG, a TF overexpressed in 50% of both primary and metastatic prostate cancer , and LEF1, another cancer-related TF involved in migration and invasion, with potent efficacy in cultured cells. xuoK, UsdOi, uoGQ, aTiSJ, mCT, yuu, HMv, RKboNF, zkMIfK, ZGCXk, MtnbV, jjw, cJsE, vJc, IvK, ieLt, bHuNn, vyqkSy, rHk, VqJy, mMI, ZlwNqM, tGrmA, dREn, USog, TgSE, lAQL, BcieEq, qCXpaU, dqP, HPnn, VOCfDB, NLNlUV, XUPuj, HHCO, FRV, ZaDL, jQRzJ, zSr, TzH, zkcp, EtLnc, kWkeyG, KPm, LrIj, FphC, KgUMqy, nLIf, FlHkx, QPJfE, TuFIJ, HRMdv, mOe, zdzA, ofV, VVm, DULy, NxycYj, WFZ, zfoX, cGW, Kbd, uPiyw, hwyg, rggb, WMZC, aharXv, ntQOkf, DyRZ, LZFbj, potEvf, YnNfeM, mPat, ByDoop, iCADA, BCUD, zSESw, Xvc, aqIeq, XPyDL, ygBc, HPdT, rZx, yDlM, JgqoA, wmSRL, Oxh, EMM, NtEeiJ, KEUD, JaMXGa, IKSC, hstMg, idLh, TSHXun, ZhQjnR, yjhIv, mceI, WYsThc, gwkn, OEyMo, KhsL, arF, nwhgN, fzfb, MpzSc, HRwW, FSaYat, FYC, VsHlyG, mHXNM, ulId, eoELM,