Ephrin A4-Ephrin Receptor A10 Signaling Promotes Cell Migration
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Ephrin-A4/Fc Chimera Human
EPHRIN-A4 EXTRACELLULAR DOMAIN/FC CHIMERA Human, Recombinant Expressed in NSO mouse myeloma cells Product Number E 0403 Storage Temperature –20 °C Synonyms: LERK-4; EFL-4 Reagents Recombinant human Ephrin-A4 extracellular domain/Fc Product Description chimera is supplied as approximately 200 mg of protein Recombinant human Ephrin-A4 extracellular domain/Fc lyophilized from a sterile-filtered phosphate-buffered chimera consists of amino acid residues 1-171 saline (PBS) solution. 1 (extracellular domain of human Ephrin-A4) that was fused by means of a polypeptide linker to the Fc portion Preparation Instructions of human IgG1 that is 6X histidine-tagged at the Reconstitute the vial contents with sterile PBS. carboxyl terminus. The chimeric protein is expressed in Stock solution concentration should be no less than a mouse myeloma cell line, NSO. Recombinant Ephrin 100 µg/ml. A4 is a disulfide-linked homodimer. The amino terminus is Leu 26 determined by N-terminal Storage/Stability sequencing. The calculated molecular mass of the Lyophilized samples are stable for greater than six reduced protein is approximately 43.7 kDa, but as a months at –20 °C. Upon reconstitution, store at 2-4 °C result of glycosylation, the recombinant Ephrin-A4/Fc for up to one month. For extended storage, store in migrates as an approximately 50 kDa protein on working aliquots at –20 °C. Repeated freeze-thaw reducing SDS -PAGE. cycles should be avoided. Do not store in frost-free freezer. The Ephrin ligand family, of which Ephrin-A4 is a member, binds members of the Eph receptor family. All Product Profile ligands share a conserved extracellular sequence, Identity of Ephrin-A4/Fc was determined by western thought to correspond to the receptor binding domain. -
Epha Receptors and Ephrin-A Ligands Are Upregulated by Monocytic
Mukai et al. BMC Cell Biology (2017) 18:28 DOI 10.1186/s12860-017-0144-x RESEARCHARTICLE Open Access EphA receptors and ephrin-A ligands are upregulated by monocytic differentiation/ maturation and promote cell adhesion and protrusion formation in HL60 monocytes Midori Mukai, Norihiko Suruga, Noritaka Saeki and Kazushige Ogawa* Abstract Background: Eph signaling is known to induce contrasting cell behaviors such as promoting and inhibiting cell adhesion/ spreading by altering F-actin organization and influencing integrin activities. We have previously demonstrated that EphA2 stimulation by ephrin-A1 promotes cell adhesion through interaction with integrins and integrin ligands in two monocyte/ macrophage cell lines. Although mature mononuclear leukocytes express several members of the EphA/ephrin-A subclass, their expression has not been examined in monocytes undergoing during differentiation and maturation. Results: Using RT-PCR, we have shown that EphA2, ephrin-A1, and ephrin-A2 expression was upregulated in murine bone marrow mononuclear cells during monocyte maturation. Moreover, EphA2 and EphA4 expression was induced, and ephrin-A4 expression was upregulated, in a human promyelocytic leukemia cell line, HL60, along with monocyte differentiation toward the classical CD14++CD16− monocyte subset. Using RT-PCR and flow cytometry, we have also shown that expression levels of αL, αM, αX, and β2 integrin subunits were upregulated in HL60 cells along with monocyte differentiation while those of α4, α5, α6, and β1 subunits were unchanged. Using a cell attachment stripe assay, we have shown that stimulation by EphA as well as ephrin-A, likely promoted adhesion to an integrin ligand- coated surface in HL60 monocytes. Moreover, EphA and ephrin-A stimulation likely promoted the formation of protrusions in HL60 monocytes. -
Engrailed and Fgf8 Act Synergistically to Maintain the Boundary Between Diencephalon and Mesencephalon Scholpp, S., Lohs, C
5293 Erratum Engrailed and Fgf8 act synergistically to maintain the boundary between diencephalon and mesencephalon Scholpp, S., Lohs, C. and Brand, M. Development 130, 4881-4893. An error in this article was not corrected before going to press. Throughout the paper, efna4 should be read as EphA4, as the authors are referring to the gene encoding the ephrin A4 receptor (EphA4) and not to that encoding its ligand ephrin A4. We apologise to the authors and readers for this mistake. Research article 4881 Engrailed and Fgf8 act synergistically to maintain the boundary between diencephalon and mesencephalon Steffen Scholpp, Claudia Lohs and Michael Brand* Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG), Dresden, Germany Department of Genetics, University of Dresden (TU), Pfotenhauer Strasse 108, 01307 Dresden, Germany *Author for correspondence (e-mail: [email protected]) Accepted 23 June 2003 Development 130, 4881-4893 © 2003 The Company of Biologists Ltd doi:10.1242/dev.00683 Summary Specification of the forebrain, midbrain and hindbrain However, a patch of midbrain tissue remains between the primordia occurs during gastrulation in response to signals forebrain and the hindbrain primordia in such embryos. that pattern the gastrula embryo. Following establishment This suggests that an additional factor maintains midbrain of the primordia, each brain part is thought to develop cell fate. We find that Fgf8 is a candidate for this signal, as largely independently from the others under the influence it is both necessary and sufficient to repress pax6.1 and of local organizing centers like the midbrain-hindbrain hence to shift the DMB anteriorly independently of the boundary (MHB, or isthmic) organizer. -
Supplemental Material 1
Supplemental Fig. 1 12 1mg/ml A1AT p<0.001 10 8 p=0.004 6 p=0.029 (fold change) 4 mRNA ofmRNA angptl4 at h 1 2 0 Control Prolastin Aralast A1AT Sigma Supplemental Figure 1. Adherent PBMCs were incubated with 1 mg/ml A1AT from different sources (Prolastin, Grifols; Aralast, Baxter; or Sigma Aldrich) for 1h and mRNA levels of angplt4 were determined by RT-PCR. Bars represent the mean ± SD of four experiments. Supplemental Fig. 2 12 8 h p=0.005 10 8 6 mRNA angplt4 mRNA change Fold 4 2 0 Control A1AT 0.5 mg/ml Supplemental Figure 2. HMVEC-L were incubated with 0.5 mg/ml A1AT (Calbiochem) for 8 h and mRNA levels of angptl4 were determined by RT-PCR. Bars represent the mean ± SEM of three experiments. Supplemental Table I. Influence of A1AT (1 mg/ml, 8 h) on expression of angiogenesis genesa in HMVEC-L Unigene Symbol Description Gname fold changeb Hs.525622 AKT1 V-akt murine thymoma viral AKT/MGC99656/PKB/PKB- 0,94 0,14 oncogene homolog 1 ALPHA/PRKBA/RAC/RAC- ALPHA Hs.369675 ANGPT1 Angiopoietin 1 AGP1/AGPT/ANG1 3,61 5,11 Hs.583870 ANGPT2 Angiopoietin 2 AGPT2/ANG2 0,98 0,01 Hs.209153 ANGPTL3 Angiopoietin-like 3 ANGPT5/FHBL2 1,10 0,59 Hs.9613 ANGPTL4 Angiopoietin-like 4 ANGPTL2/ARP4/FIAF/HFAR 5,79 1,66 P/NL2/PGAR/pp1158 Hs.1239 ANPEP Alanyl (membrane) APN/CD13/GP150/LAP1/P150 1,03 0,01 aminopeptidase /PEPN Hs.194654 BAI1 Brain-specific angiogenesis FLJ41988/GDAIF 0,89 0,26 inhibitor 1 Hs.54460 CCL11 Chemokine (C-C motif) ligand 11 MGC22554/SCYA11 1,04 0,15 Hs.303649 CCL2 Chemokine (C-C motif) ligand 2 GDCF- 1,09 0,14 2/HC11/HSMCR30/MCAF/M CP- -
Mtorc1-Independent Autophagy Regulates Receptor Tyrosine Kinase Phosphorylation in Colorectal Cancer Cells Via an Mtorc2-Mediated Mechanism
Cell Death and Differentiation (2017) 24, 1045–1062 & 2017 Macmillan Publishers Limited, part of Springer Nature. All rights reserved 1350-9047/17 www.nature.com/cdd mTORC1-independent autophagy regulates receptor tyrosine kinase phosphorylation in colorectal cancer cells via an mTORC2-mediated mechanism Aikaterini Lampada1,2, James O’Prey3, Gyorgy Szabadkai4, Kevin M Ryan3, Daniel Hochhauser*,2,5 and Paolo Salomoni*,1,5 The intracellular autophagic degradative pathway can have a tumour suppressive or tumour-promoting role depending on the stage of tumour development. Upon starvation or targeting of oncogenic receptor tyrosine kinases (RTKs), autophagy is activated owing to the inhibition of PI3K/AKT/mTORC1 signalling pathway and promotes survival, suggesting that autophagy is a relevant therapeutic target in these settings. However, the role of autophagy in cancer cells where the PI3K/AKT/mTORC1 pathway is constitutively active remains partially understood. Here we report a role for mTORC1-independent basal autophagy in regulation of RTK activation and cell migration in colorectal cancer (CRC) cells. PI3K and RAS-mutant CRC cells display basal autophagy levels despite constitutive mTORC1 signalling, but fail to increase autophagic flux upon RTK inhibition. Inhibition of basal autophagy via knockdown of ATG7 or ATG5 leads to decreased phosphorylation of several RTKs, in particular c-MET. Internalised c-MET colocalised with LAMP1-negative, LC3-positive vesicles. Finally, autophagy regulates c-MET phosphorylation via an mTORC2- dependent -
Ephrin-A Binding and Epha Receptor Expression Delineate the Matrix Compartment of the Striatum
The Journal of Neuroscience, June 15, 1999, 19(12):4962–4971 Ephrin-A Binding and EphA Receptor Expression Delineate the Matrix Compartment of the Striatum L. Scott Janis, Robert M. Cassidy, and Lawrence F. Kromer Department of Cell Biology and Interdisciplinary Program in Neuroscience, Georgetown University Medical Center, Washington, DC 20007 The striatum integrates limbic and neocortical inputs to regulate matrix neurons. In situ hybridization for EphA RTKs reveals that sensorimotor and psychomotor behaviors. This function is de- the two different ligand binding patterns strictly match pendent on the segregation of striatal projection neurons into the mRNA expression patterns of EphA4 and EphA7. anatomical and functional components, such as the striosome Ligand–receptor binding assays indicate that ephrin-A1 and and matrix compartments. In the present study the association ephrin-A4 selectively bind EphA4 but not EphA7 in the lysates of ephrin-A cell surface ligands and EphA receptor tyrosine of striatal tissue. Conversely, ephrin-A2, ephrin-A3, and kinases (RTKs) with the organization of these compartments ephrin-A5 bind EphA7 but not EphA4. These observations im- was determined in postnatal rats. Ephrin-A1 and ephrin-A4 plicate selective interactions between ephrin-A molecules and selectively bind to EphA receptors on neurons restricted to the EphA RTKs as potential mechanisms for regulating the com- matrix compartment. Binding is absent from the striosomes, partmental organization of the striatum. which were identified by m-opioid -
Development and Validation of a Protein-Based Risk Score for Cardiovascular Outcomes Among Patients with Stable Coronary Heart Disease
Supplementary Online Content Ganz P, Heidecker B, Hveem K, et al. Development and validation of a protein-based risk score for cardiovascular outcomes among patients with stable coronary heart disease. JAMA. doi: 10.1001/jama.2016.5951 eTable 1. List of 1130 Proteins Measured by Somalogic’s Modified Aptamer-Based Proteomic Assay eTable 2. Coefficients for Weibull Recalibration Model Applied to 9-Protein Model eFigure 1. Median Protein Levels in Derivation and Validation Cohort eTable 3. Coefficients for the Recalibration Model Applied to Refit Framingham eFigure 2. Calibration Plots for the Refit Framingham Model eTable 4. List of 200 Proteins Associated With the Risk of MI, Stroke, Heart Failure, and Death eFigure 3. Hazard Ratios of Lasso Selected Proteins for Primary End Point of MI, Stroke, Heart Failure, and Death eFigure 4. 9-Protein Prognostic Model Hazard Ratios Adjusted for Framingham Variables eFigure 5. 9-Protein Risk Scores by Event Type This supplementary material has been provided by the authors to give readers additional information about their work. Downloaded From: https://jamanetwork.com/ on 10/02/2021 Supplemental Material Table of Contents 1 Study Design and Data Processing ......................................................................................................... 3 2 Table of 1130 Proteins Measured .......................................................................................................... 4 3 Variable Selection and Statistical Modeling ........................................................................................ -
Rabbit Anti-Ephrin-A1 Rabbit Anti-Ephrin-A1
Qty: 100 µg/400 µl Rabbit anti-ephrin-A1 Catalog No. 34-3300 Lot No. See product label Rabbit anti-ephrin-A1 FORM This polyclonal antibody is supplied as a 400 µl aliquot at a concentration of 0.25 mg/ml in phosphate buffered saline (pH 7.4) containing 0.1% sodium azide. The antibody is epitope-affinity-purified from rabbit antiserum. PAD: ZMD.39 IMMUNOGEN Synthetic peptide derived from the C-terminal end of mouse ephrin-A1 protein. SPECIFICITY This antibody is specific for the ephrin-A1 protein. REACTIVITY Reactivity is confirmed with a chimeric protein consisting of the extracellular domain of mouse ephrin-A1 and the Fc region of human IgG1. Cross-reactivity with human ephrin-A1 is confirmed with IHC experiments on human tissue sections and this reactivity was expected because of the 85% shared amino acid identity in the extracellular domain. Sample Western Immunoprecipitation Immunohistochemistry Blotting (FFPE) Mouse +++ +++ NT Human NT NT ++ (Excellent +++, Good++, Poor +, No reactivity 0, Not tested NT) USAGE Working concentrations for specific applications should be determined by the investigator. Appropriate concentrations will be affected by several factors, including secondary antibody affinity, antigen concentration, sensitivity of detection method, temperature and length of incubations, etc. The suitability of this antibody for applications other than those listed below has not been determined. The following concentration ranges are recommended starting points for this product. Western Blotting: 1-5 µg/mL Immunoprecipitation: 5-10 µg/ IP reaction Immunohistochemistry: 4-10 µg/mL Please note that immunohistochemical assays were optimized on formalin-fixed, paraffin-embedded tissue sections and Heat Induced Epitope Retrieval (HIER) with EDTA, pH 8.0 was required for optimal staining. -
Protein Tyrosine Kinases: Their Roles and Their Targeting in Leukemia
cancers Review Protein Tyrosine Kinases: Their Roles and Their Targeting in Leukemia Kalpana K. Bhanumathy 1,*, Amrutha Balagopal 1, Frederick S. Vizeacoumar 2 , Franco J. Vizeacoumar 1,3, Andrew Freywald 2 and Vincenzo Giambra 4,* 1 Division of Oncology, College of Medicine, University of Saskatchewan, Saskatoon, SK S7N 5E5, Canada; [email protected] (A.B.); [email protected] (F.J.V.) 2 Department of Pathology and Laboratory Medicine, College of Medicine, University of Saskatchewan, Saskatoon, SK S7N 5E5, Canada; [email protected] (F.S.V.); [email protected] (A.F.) 3 Cancer Research Department, Saskatchewan Cancer Agency, 107 Wiggins Road, Saskatoon, SK S7N 5E5, Canada 4 Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapies (ISBReMIT), Fondazione IRCCS Casa Sollievo della Sofferenza, 71013 San Giovanni Rotondo, FG, Italy * Correspondence: [email protected] (K.K.B.); [email protected] (V.G.); Tel.: +1-(306)-716-7456 (K.K.B.); +39-0882-416574 (V.G.) Simple Summary: Protein phosphorylation is a key regulatory mechanism that controls a wide variety of cellular responses. This process is catalysed by the members of the protein kinase su- perfamily that are classified into two main families based on their ability to phosphorylate either tyrosine or serine and threonine residues in their substrates. Massive research efforts have been invested in dissecting the functions of tyrosine kinases, revealing their importance in the initiation and progression of human malignancies. Based on these investigations, numerous tyrosine kinase inhibitors have been included in clinical protocols and proved to be effective in targeted therapies for various haematological malignancies. -
Molecular Regulation of Visual System Development: More Than Meets the Eye
Downloaded from genesdev.cshlp.org on September 30, 2021 - Published by Cold Spring Harbor Laboratory Press REVIEW Molecular regulation of visual system development: more than meets the eye Takayuki Harada,1,2 Chikako Harada,1,2 and Luis F. Parada1,3 1Department of Developmental Biology and Kent Waldrep Foundation Center for Basic Neuroscience Research on Nerve Growth and Regeneration, University of Texas Southwestern Medical Center, Dallas, Texas 75235, USA; 2Department of Molecular Neurobiology, Tokyo Metropolitan Institute for Neuroscience, Fuchu, Tokyo 183-8526, Japan Vertebrate eye development has been an excellent model toderm, intercalating mesoderm, surface ectoderm, and system to investigate basic concepts of developmental neural crest (Fig. 1). The neuroectoderm differentiates biology ranging from mechanisms of tissue induction to into the retina, iris, and optic nerve; the surface ecto- the complex patterning and bidimensional orientation of derm gives rise to lens and corneal epithelium; the me- the highly specialized retina. Recent advances have shed soderm differentiates into the extraocular muscles and light on the interplay between numerous transcriptional the fibrous and vascular coats of the eye; and neural crest networks and growth factors that are involved in the cells become the corneal stroma sclera and corneal en- specific stages of retinogenesis, optic nerve formation, dothelium. The vertebrate eye originates from bilateral and topographic mapping. In this review, we summarize telencephalic optic grooves. In humans, optic vesicles this recent progress on the molecular mechanisms un- emerge at the end of the fourth week of development and derlying the development of the eye, visual system, and soon thereafter contact the surface ectoderm to induce embryonic tumors that arise in the optic system. -
ALL1 Fusion Proteins Induce Deregulation of Epha7 and ERK Phosphorylation in Human Acute Leukemias
ALL1 fusion proteins induce deregulation of EphA7 and ERK phosphorylation in human acute leukemias Hiroshi Nakanishi*, Tatsuya Nakamura*, Eli Canaani†, and Carlo M. Croce*‡ *Department of Molecular Virology, Immunology, and Medical Genetics and Comprehensive Cancer Center, Ohio State University, Columbus, OH 43210; and †Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot 76100, Israel Edited by Janet D. Rowley, University of Chicago Medical Center, Chicago, IL, and approved July 25, 2007 (received for review April 6, 2007) Erythropoietin-producing hepatoma-amplified sequence (Eph) re- studies revealed deregulation of some of the Eph/ephrin genes in ceptor tyrosine kinases and their cell-surface-bound ligands, the human malignancies, including up-regulation of ephrin-A1 or -B2 ephrins, function as a unique signaling system triggered by cell- in melanoma (2, 3), up-regulation of EphB2 in stomach cancer to-cell interaction and have been shown to mediate neurodevel- (4) and in breast cancer (5), and up-regulation of EphA2 in opmental processes. In addition, recent studies showed deregula- prostate (6), breast (7), and esophageal cancers (8), some of tion of some of Eph/ephrin genes in human malignancies, which were shown to be associated with tumor invasion or tumor suggesting the involvement of this signaling pathway in tumori- metastasis and therefore associated with poor prognosis. Con- genesis. The ALL1 (also termed MLL) gene on human chromosome versely, mutational inactivation of EphB2 was detected in pros- 11q23 was isolated by virtue of its involvement in recurrent tate (9) and colon cancers (10), suggesting tumor suppressor chromosome translocations associated with acute leukemias with function of this Eph receptor in the relevant tumors. -
Epha3 Function Is Regulated by Multiple
EPHA3 FUNCTION IS REGULA TED BY MULTIPLE PHOSPHOTYROSINE RESIDUES 2 1 Guanfang Shi\ Gang Yue , Mike Sheng\ Suzie Chen\ and Renping Zhou IDepartment of Chemical Biology, Ernest Mario School of Pharmacy, Rutgers University, Piscataway, NJ 08854; 2Department of Oral Biology, New Jersey Dental School, University of Medicine & Dentistry of New Jersey, Newark, NJ 07101. Address correspondence to: Renping Zhou, Ph. D. Phone: (732) 445-3400 ext. 264; Fax: (732) 445-0687; E-mail: rzhou @rci.rutgers.edu EphA3, a member of the Eph family of tyrosine carcinoma was significantly correlated with kinase receptors, has been implicated in a regional lymph node metastasis (5). variety of tumors including melanoma, Overexpression of ephrin-B2, a ligand for EphB glioblastoma, pancreatic carcinoma, lung and receptors, in B 16 melanoma cells enhanced colon cancers. The loss of EphA3 function has integrin-mediated ECM-attachment and migration been associated with colorectal cancer. To (6). EphB2 positively regulates glioma cell identify potential roles in tumor formation, we adhesion, growth, and invasion via R-Ras signaling examined effects of EphA3 expression and (7). In contrast to promote tumor metastasis and activation on melanoma cell migration, and cancer cell line migration, Eph receptors may also identified key signal transducer docking sites of regulate the cell motility negatively. Hafner and the receptor. This analysis showed that the coworkers compared EphB6 mRNA level in human melanoma cell line WMl15 lacks benign nevi, primary melanomas, and metastatic EphA3 expression, while two other melanoma melanomas, and found a progressive and cell lines, WM239A and C8161, both retain significant reduction of EphB6 mRNA expression expression of the receptor.