DOCSLIB.ORG

REGULATION of APICAL POLARITY COMPLEXES by Jay Nicholas

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
REGULATION of APICAL POLARITY COMPLEXES by Jay Nicholas REGULATION OF APICAL POLARITY COMPLEXES By Jay Nicholas Pieczynski A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy (Biological Chemistry) in the University of Michigan 2010 Doctoral Committee: Professor Benjamin L. Margolis, Chair Professor Eric R. Fearon Professor Robert S. Fuller Assistant Professor Daniel A. Bochar Assistant Professor Hisashi Umemori Acknowledgements I would like to thank my mentor Dr. Ben Margolis for his guidance and support throughout the process of earning my doctorate degree. He has taught me that being a scientist means being your own toughest critic and to always remember that your work represents who you are and what you have accomplished. I would also like to thank the current and former members of the Margolis laboratory who made my time in the lab very enjoyable. Each person past and present has brought their own personality to the lab and contributed to the success of the group. It has been an honor to be associated with truly gifted scientists, yet even better people. The work contained within this thesis is also dedicated to my former mentors especially Michael Baxter and Dr. Todd Weaver. Michael Baxter was a former teacher of mine growing up in Wisconsin and recognized my potential as a scientist at a very early age. He taught me that science is not about how intelligent you are, but asking the write questions. Dr. Todd Weaver was also very instrumental in my scientific career, allowing me to begin my independent research career during the first years of college. Dr. Weaver recognized my desire to always know more, and challenged me to work outside the classroom in his laboratory for most of my undergraduate career; an experience that I am forever indebted to. Finally, I would like to thank my friends and family. I am blessed to have some of the best friends that one could ask for and my success can only be attributed to having ii people back me up all the way. My brother Tad has been my best friend my entire life and always encouraged me to be the best at whatever I do. My parents, Guy and Liz, have always been there for me especially when times got tough. They taught me that nothing in life is free and that hard work always eventually pays off. I am truly lucky to have two of the best parents who always wanted more for their children than they had. iii Preface Chapter 1 will be submitted as part of an invited review for the American Journal of Physiology Renal Physiology. Chapter 2 was published in December of 2006 in the Journal of Biological Chemistry and contains work completed by Dr. Samuel Straight and Jay Pieczynski. Chapter 3 contains data that will be submitted as a manuscript to Experimental Cell Research and is collaboration between Jay Pieczynski, Dr. Sanjeevkumar Patel, and Dr. Peter Arvan. Chapter 4 contains work that will be published in a future manuscript. Appendix A contains the full data sets of the microarray analysis completed in Chapter 3. Appendix B contains data that was published in Schlüter et al. in Molecular and Cellular Biology in September of 2009 and unpublished data related to that manuscript. iv Table of Contents Acknowledgements............................................................................................................ii Preface ...............................................................................................................................iv List of Figures ................................................................................................................ viii List of Tables......................................................................................................................x List of Appendices ............................................................................................................xi Chapter 1: Development and Maintenance of Renal Epithelial Cell Polarity.............1 1.1 Introduction ...........................................................................................................1 1.2 Molecular Interactions Governing the Establishment of Mammalian Epithelial Cell Polarity.................................................................................................................4 1.3 Mammalian Polarity Complexes and Their Interactions.......................................7 1.3.1 The Par Complex Overview ...............................................................7 1.3.2 Par3...................................................................................................10 1.3.3 Par6...................................................................................................15 1.3.4 aPKCζ ..............................................................................................18 1.3.5 The Scribble Complex Overview .....................................................20 1.3.6 Scribble.............................................................................................20 1.3.7 Lgl ....................................................................................................22 1.3.8 Dlg ....................................................................................................24 1.3.9 Crumbs complex Overview..............................................................27 1.3.10 PALS1 ............................................................................................27 1.3.11 PATJ and MUPP1 ..........................................................................29 1.3.12 Crumbs3 .........................................................................................32 1.4 Molecular Mechanisms Establishing Renal Epithelial Cell Polarity ..................35 1.4.1 Establishing Renal Polarity: MET vs. EMT.....................................35 1.4.2 Regulation and Maintenance of Renal Epithelial Polarity: A Trail of Crumbs ......................................................................................................37 1.4.3 Renal Apico-Basal Epithelial Polarity in Context: A Link to Nephropathies, Planar Cell Polarity, and Renal Cilia ...............................42 v 1.5 Concluding Remarks ...........................................................................................45 Chapter 2: Mammalian Lin-7 Stabilizes Polarity Complex Proteins.........................47 2.1 Introduction .........................................................................................................47 2.2 Materials and Methods ........................................................................................49 2.2.1 DNA Constructs ...............................................................................49 2.2.2 Cell Culture and Transfection ..........................................................50 2.2.3 Calcium Switch Assay and Transepithelial Electrical Resistance Measurement .............................................................................................51 2.2.4 Antibodies ........................................................................................52 2.2.5 Immunoprecipitation and Immunoblotting ......................................53 2.2.6 CRB3 Peptide Beads ........................................................................54 2.2.7 Pulse-Chase Labeling .......................................................................54 2.2.8 Immunostaining and Confocal Microscopy .....................................55 2.2.9 Quantitative Real-time PCR.............................................................56 2.3 Results .................................................................................................................57 2.3.1 Lin-7 shRNA affects the formation of tight junctions .....................57 2.3.2 Lin-7 shRNA affects expression of MAGUK family members.......61 2.3.3 The L27 domain of Lin-7C is essential for correct tight junction formation ...................................................................................................63 2.3.4 Reexpression of PALS1 can reverse the tight junction defect seen in Lin-7 knockdown cells ..............................................................................66 2.3.5 Lin-7 expression affects stability of PALS1 in polarized MDCKII cells............................................................................................................73 2.4 Discussion ...........................................................................................................75 Chapter 3: Slug Represses Crumbs3 in Epithelial Cells ..............................................78 3.1 Introduction .........................................................................................................78 3.2 Materials and Methods ........................................................................................81 3.2.1 Cell lines...........................................................................................81 3.2.2 DNA Constructs ...............................................................................82 3.2.3 Antibodies and Immunostaining ......................................................83 3.2.4 Microarray and Statistics..................................................................84 3.2.5 qRT-PCR..........................................................................................84 3.2.6 ChIP Assays .....................................................................................85 3.3 Results .................................................................................................................87
Load more

Recommended publications
  • Association Analyses of Known Genetic Variants with Gene
    ASSOCIATION ANALYSES OF KNOWN GENETIC VARIANTS WITH GENE EXPRESSION IN BRAIN by Viktoriya Strumba A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy (Bioinformatics) in The University of Michigan 2009 Doctoral Committee: Professor Margit Burmeister, Chair Professor Huda Akil Professor Brian D. Athey Assistant Professor Zhaohui S. Qin Research Statistician Thomas Blackwell To Sam and Valentina Dmitriy and Elizabeth ii ACKNOWLEDGEMENTS I would like to thank my advisor Professor Margit Burmeister, who tirelessly guided me though seemingly impassable corridors of graduate work. Throughout my thesis writing period she provided sound advice, encouragement and inspiration. Leading by example, her enthusiasm and dedication have been instrumental in my path to becoming a better scientist. I also would like to thank my co-advisor Tom Blackwell. His careful prodding always kept me on my toes and looking for answers, which taught me the depth of careful statistical analysis. His diligence and dedication have been irreplaceable in most difficult of projects. I also would like to thank my other committee members: Huda Akil, Brian Athey and Steve Qin as well as David States. You did not make it easy for me, but I thank you for believing and not giving up. Huda’s eloquence in every subject matter she explained have been particularly inspiring, while both Huda’s and Brian’s valuable advice made the completion of this dissertation possible. I would also like to thank all the members of the Burmeister lab, both past and present: Sandra Villafuerte, Kristine Ito, Cindy Schoen, Karen Majczenko, Ellen Schmidt, Randi Burns, Gang Su, Nan Xiang and Ana Progovac.
    [Show full text]
  • A Rare Missense Mutation in GJB3 (Cx31g45e) Is Associated with a Unique Cellular Phenotype Resulting in Necrotic Cell Death Easton, J
    University of Dundee A rare missense mutation in GJB3 (Cx31G45E) is associated with a unique cellular phenotype resulting in necrotic cell death Easton, J. A.; Alboulshi, A. K.; Kamps, M. A. F.; Brouns, G. H.; Broers, M. R.; Coull, B. J.; Oji, V.; van Geel, M.; van Steensel, M. A. M.; Martin, P. E. Published in: Experimental Dermatology DOI: 10.1111/exd.13542 Publication date: 2018 Document Version Peer reviewed version Link to publication in Discovery Research Portal Citation for published version (APA): Easton, J. A., Alboulshi, A. K., Kamps, M. A. F., Brouns, G. H., Broers, M. R., Coull, B. J., ... Martin, P. E. (2018). A rare missense mutation in GJB3 (Cx31G45E) is associated with a unique cellular phenotype resulting in necrotic cell death. Experimental Dermatology. https://doi.org/10.1111/exd.13542 General rights Copyright and moral rights for the publications made accessible in Discovery Research Portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. • Users may download and print one copy of any publication from Discovery Research Portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain. • You may freely distribute the URL identifying the publication in the public portal. Take down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim.
    [Show full text]
  • Supplemental Figure 1. Vimentin
    Double mutant specific genes Transcript gene_assignment Gene Symbol RefSeq FDR Fold- FDR Fold- FDR Fold- ID (single vs. Change (double Change (double Change wt) (single vs. wt) (double vs. single) (double vs. wt) vs. wt) vs. single) 10485013 BC085239 // 1110051M20Rik // RIKEN cDNA 1110051M20 gene // 2 E1 // 228356 /// NM 1110051M20Ri BC085239 0.164013 -1.38517 0.0345128 -2.24228 0.154535 -1.61877 k 10358717 NM_197990 // 1700025G04Rik // RIKEN cDNA 1700025G04 gene // 1 G2 // 69399 /// BC 1700025G04Rik NM_197990 0.142593 -1.37878 0.0212926 -3.13385 0.093068 -2.27291 10358713 NM_197990 // 1700025G04Rik // RIKEN cDNA 1700025G04 gene // 1 G2 // 69399 1700025G04Rik NM_197990 0.0655213 -1.71563 0.0222468 -2.32498 0.166843 -1.35517 10481312 NM_027283 // 1700026L06Rik // RIKEN cDNA 1700026L06 gene // 2 A3 // 69987 /// EN 1700026L06Rik NM_027283 0.0503754 -1.46385 0.0140999 -2.19537 0.0825609 -1.49972 10351465 BC150846 // 1700084C01Rik // RIKEN cDNA 1700084C01 gene // 1 H3 // 78465 /// NM_ 1700084C01Rik BC150846 0.107391 -1.5916 0.0385418 -2.05801 0.295457 -1.29305 10569654 AK007416 // 1810010D01Rik // RIKEN cDNA 1810010D01 gene // 7 F5 // 381935 /// XR 1810010D01Rik AK007416 0.145576 1.69432 0.0476957 2.51662 0.288571 1.48533 10508883 NM_001083916 // 1810019J16Rik // RIKEN cDNA 1810019J16 gene // 4 D2.3 // 69073 / 1810019J16Rik NM_001083916 0.0533206 1.57139 0.0145433 2.56417 0.0836674 1.63179 10585282 ENSMUST00000050829 // 2010007H06Rik // RIKEN cDNA 2010007H06 gene // --- // 6984 2010007H06Rik ENSMUST00000050829 0.129914 -1.71998 0.0434862 -2.51672
    [Show full text]
  • Epithelia and Gastrointestinal Function
    CHAPTER 18 Epithelia and gastrointestinal function Jerrold R. Turner University of Chicago, Chicago, IL, USA Chapter menu Organization of the gut wall, 317 Epithelial barrier and disease , 326 Organization of epithelial cells and sheets, 318 Integration of mucosal function, 329 Mucosal barriers, 322 Further reading, 329 All cavities within the alimentary tract, from the small ducts An underlying layer of fibroconnective tissue called the sub- and acini of the pancreas to the gastric lumen, are lined by mucosa, which contains nerves, vessels, and lymphatics, sup- sheets of polarized epithelial cells. Common to all of these epi- ports the mucosa. The submucosa rests on the muscularis thelia is the ability to create selective barriers that separate propria, which is composed of two or three layers of smooth luminal and tissue spaces. Most epithelia are also able to direct muscle and is home to the myenteric plexus (see Chapters 1, 15, vectorial transport of solutes and solvents. These essential func- and 16). In most instances, gastrointestinal organs are encased tions are based on the structural polarity of individual cells, the by an outermost delicate layer of fibrofatty tissue, the serosa, complex organization of membranes, cell–cell and cell–substrate encircled by a continuous layer of mesothelial cells. In areas interactions, and interactions with other cell types. This chapter where no serosa exists, as in portions of the esophagus and the reviews intestinal wall structure and examines how mucosal distal colorectum, fibrofatty tissues interface with the external functions are supported by the organization of the gut and the portion of the muscularis propria.
    [Show full text]
  • Molecular Biology and Biochemistry of The
    MOLECULAR BIOLOGY AND BIOCHEMISTRY OF THE REGULATION OF HRP/TYPE III SECRETION GENES IN THE CORN PATHOGEN PANTOEA STEWARTII SUBSP. STEWARTII DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Massimo Merighi, B. S., M. S. ***** The Ohio State University 2003 Dissertation Committee: Professor David L. Coplin, Adviser Approved by Professor Brian Ahmer ______________________ Professor Dietz W. Bauer Adviser Professor Terrence L. Graham Plant Pathology Copyright by Massimo Merighi 2003 ii ABSTRACT Pantoea stewartii subsp. stewartii is a bacterial pathogen of corn. Its pathogenicity depends on the expression of a Hrp/type III protein secretion/translocation system. The regulatory region of the hrp gene cluster consists of three adjacent operons: hrpXY encodes a two- component regulatory system, consisting of the response regulator HrpY and sensor PAS-kinase HrpX; hrpS encodes an NtrC-like enhancer-binding protein; and hrpL encodes an ECF sigma factor. In this study, we used genetic and biochemical approaches to delineate the following regulatory cascade: 1) HrpY activates hrpS; 2) HrpS activates hrpL; and 3) HrpL activates secretion and effector genes with ‘Hrp-box’ promoters. This pathway responds to environmental signals and global regulators. Mutant analysis showed that HrpX is required for full virulence. Deletion of its individual PAS-sensory domains revealed that they are not redundant and each may have a different role in modulating kinase activity. HrpX probably senses an intracellular signal, possibly related to nitrogen metabolism. pH, osmolarity and nicotinic acid controlled expression of hrpS independently of HrpX/HrpY.
    [Show full text]
  • Nrf2 Modulates Host Defense During Streptococcus Pneumoniae Pneumonia in Mice
    Nrf2 Modulates Host Defense during Streptococcus pneumoniae Pneumonia in Mice This information is current as John C. Gomez, Hong Dang, Jessica R. Martin and Claire of September 28, 2021. M. Doerschuk J Immunol published online 26 August 2016 http://www.jimmunol.org/content/early/2016/08/26/jimmun ol.1600043 Downloaded from Supplementary http://www.jimmunol.org/content/suppl/2016/08/26/jimmunol.160004 Material 3.DCSupplemental http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on September 28, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2016 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published August 26, 2016, doi:10.4049/jimmunol.1600043 The Journal of Immunology Nrf2 Modulates Host Defense during Streptococcus pneumoniae Pneumonia in Mice John C. Gomez,*,† Hong Dang,†,‡ Jessica R. Martin,*,† and Claire M. Doerschuk*,†,x Nrf2 regulates the transcriptional response to oxidative stress. These studies tested the role of Nrf2 during Streptococcus pneumoniae pneumonia and identified Nrf2-dependent genes and pathways in lung tissue and in recruited neutrophils.
    [Show full text]
  • Efficacy and Mechanistic Evaluation of Tic10, a Novel Antitumor Agent
    University of Pennsylvania ScholarlyCommons Publicly Accessible Penn Dissertations 2012 Efficacy and Mechanisticv E aluation of Tic10, A Novel Antitumor Agent Joshua Edward Allen University of Pennsylvania, [email protected] Follow this and additional works at: https://repository.upenn.edu/edissertations Part of the Oncology Commons Recommended Citation Allen, Joshua Edward, "Efficacy and Mechanisticv E aluation of Tic10, A Novel Antitumor Agent" (2012). Publicly Accessible Penn Dissertations. 488. https://repository.upenn.edu/edissertations/488 This paper is posted at ScholarlyCommons. https://repository.upenn.edu/edissertations/488 For more information, please contact [email protected]. Efficacy and Mechanisticv E aluation of Tic10, A Novel Antitumor Agent Abstract TNF-related apoptosis-inducing ligand (TRAIL; Apo2L) is an endogenous protein that selectively induces apoptosis in cancer cells and is a critical effector in the immune surveillance of cancer. Recombinant TRAIL and TRAIL-agonist antibodies are in clinical trials for the treatment of solid malignancies due to the cancer-specific cytotoxicity of TRAIL. Recombinant TRAIL has a short serum half-life and both recombinant TRAIL and TRAIL receptor agonist antibodies have a limited capacity to perfuse to tissue compartments such as the brain, limiting their efficacy in certain malignancies. To overcome such limitations, we searched for small molecules capable of inducing the TRAIL gene using a high throughput luciferase reporter gene assay. We selected TRAIL-inducing compound 10 (TIC10) for further study based on its induction of TRAIL at the cell surface and its promising therapeutic index. TIC10 is a potent, stable, and orally active antitumor agent that crosses the blood-brain barrier and transcriptionally induces TRAIL and TRAIL-mediated cell death in a p53-independent manner.
    [Show full text]
  • The Mineralocorticoid Receptor Leads to Increased Expression of EGFR
    www.nature.com/scientificreports OPEN The mineralocorticoid receptor leads to increased expression of EGFR and T‑type calcium channels that support HL‑1 cell hypertrophy Katharina Stroedecke1,2, Sandra Meinel1,2, Fritz Markwardt1, Udo Kloeckner1, Nicole Straetz1, Katja Quarch1, Barbara Schreier1, Michael Kopf1, Michael Gekle1 & Claudia Grossmann1* The EGF receptor (EGFR) has been extensively studied in tumor biology and recently a role in cardiovascular pathophysiology was suggested. The mineralocorticoid receptor (MR) is an important efector of the renin–angiotensin–aldosterone‑system and elicits pathophysiological efects in the cardiovascular system; however, the underlying molecular mechanisms are unclear. Our aim was to investigate the importance of EGFR for MR‑mediated cardiovascular pathophysiology because MR is known to induce EGFR expression. We identifed a SNP within the EGFR promoter that modulates MR‑induced EGFR expression. In RNA‑sequencing and qPCR experiments in heart tissue of EGFR KO and WT mice, changes in EGFR abundance led to diferential expression of cardiac ion channels, especially of the T‑type calcium channel CACNA1H. Accordingly, CACNA1H expression was increased in WT mice after in vivo MR activation by aldosterone but not in respective EGFR KO mice. Aldosterone‑ and EGF‑responsiveness of CACNA1H expression was confrmed in HL‑1 cells by Western blot and by measuring peak current density of T‑type calcium channels. Aldosterone‑induced CACNA1H protein expression could be abrogated by the EGFR inhibitor AG1478. Furthermore, inhibition of T‑type calcium channels with mibefradil or ML218 reduced diameter, volume and BNP levels in HL‑1 cells. In conclusion the MR regulates EGFR and CACNA1H expression, which has an efect on HL‑1 cell diameter, and the extent of this regulation seems to depend on the SNP‑216 (G/T) genotype.
    [Show full text]
  • KLF2 Induced
    UvA-DARE (Digital Academic Repository) The transcription factor KLF2 in vascular biology Boon, R.A. Publication date 2008 Link to publication Citation for published version (APA): Boon, R. A. (2008). The transcription factor KLF2 in vascular biology. General rights It is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), other than for strictly personal, individual use, unless the work is under an open content license (like Creative Commons). Disclaimer/Complaints regulations If you believe that digital publication of certain material infringes any of your rights or (privacy) interests, please let the Library know, stating your reasons. In case of a legitimate complaint, the Library will make the material inaccessible and/or remove it from the website. Please Ask the Library: https://uba.uva.nl/en/contact, or a letter to: Library of the University of Amsterdam, Secretariat, Singel 425, 1012 WP Amsterdam, The Netherlands. You will be contacted as soon as possible. UvA-DARE is a service provided by the library of the University of Amsterdam (https://dare.uva.nl) Download date:23 Sep 2021 Supplementary data: Genes induced by KLF2 Dekker et al. LocusLink Accession Gene Sequence Description Fold p-value ID number symbol change (FDR) 6654 AK022099 SOS1 cDNA FLJ12037 fis, clone HEMBB1001921. 100.00 5.9E-09 56999 AF086069 ADAMTS9 full length insert cDNA clone YZ35C05. 100.00 1.2E-09 6672 AF085934 SP100 full length insert cDNA clone YR57D07. 100.00 6.7E-13 9031 AF132602 BAZ1B Williams Syndrome critical region WS25 mRNA, partial sequence.
    [Show full text]
  • GJA4/Connexin 37 Mutations Correlate with Secondary Lymphedema Following Surgery in Breast Cancer Patients
    biomedicines Article GJA4/Connexin 37 Mutations Correlate with Secondary Lymphedema Following Surgery in Breast Cancer Patients Mahrooyeh Hadizadeh 1,2, Seiied Mojtaba Mohaddes Ardebili 1, Mansoor Salehi 2, Chris Young 3, Fariborz Mokarian 4, James McClellan 5, Qin Xu 6, Mohammad Kazemi 2, Elham Moazam 4, Behzad Mahaki 7 ID and Maziar Ashrafian Bonab 8,* 1 Department of Medical Genetics, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz 5166614766, Iran; [email protected] (M.H.); [email protected] (S.M.M.A.) 2 Department of Genetics and Molecular Biology, Isfahan University of Medical Sciences, Isfahan 81746753461, Iran; [email protected] (M.S.); [email protected] (M.K.) 3 School of Allied Health Sciences, Faculty of Health and Life Sciences, De Montfort University, Leicester LE1 9BH, UK; [email protected] 4 Cancer Prevention Research Centre, Isfahan University of Medical Sciences, Isfahan 8184917911, Iran; [email protected] (F.M.); [email protected] (E.M.) 5 School of Biological Sciences, University of Portsmouth, Portsmouth PO1 2DY, UK; [email protected] 6 School of Pharmacy, Faculty of Health and Life Sciences, De Montfort University, Leicester LE1 9BH, UK; [email protected] 7 Department of Occupational Health Engineering, School of Health, Isfahan University of Medical Sciences, Isfahan 8174673461, Iran; [email protected] 8 Department of Biological Sciences, University of Chester, Chester CH1 4BJ, UK * Correspondence: [email protected]; Tel.: +44-(0)1244-513-056 Received: 31 December 2017; Accepted: 13 February 2018; Published: 22 February 2018 Abstract: Lymphedema is a condition resulting from mutations in various genes essential for lymphatic development and function, which leads to obstruction of the lymphatic system.
    [Show full text]
  • Table 2. Significant
    Table 2. Significant (Q < 0.05 and |d | > 0.5) transcripts from the meta-analysis Gene Chr Mb Gene Name Affy ProbeSet cDNA_IDs d HAP/LAP d HAP/LAP d d IS Average d Ztest P values Q-value Symbol ID (study #5) 1 2 STS B2m 2 122 beta-2 microglobulin 1452428_a_at AI848245 1.75334941 4 3.2 4 3.2316485 1.07398E-09 5.69E-08 Man2b1 8 84.4 mannosidase 2, alpha B1 1416340_a_at H4049B01 3.75722111 3.87309653 2.1 1.6 2.84852656 5.32443E-07 1.58E-05 1110032A03Rik 9 50.9 RIKEN cDNA 1110032A03 gene 1417211_a_at H4035E05 4 1.66015788 4 1.7 2.82772795 2.94266E-05 0.000527 NA 9 48.5 --- 1456111_at 3.43701477 1.85785922 4 2 2.8237185 9.97969E-08 3.48E-06 Scn4b 9 45.3 Sodium channel, type IV, beta 1434008_at AI844796 3.79536664 1.63774235 3.3 2.3 2.75319499 1.48057E-08 6.21E-07 polypeptide Gadd45gip1 8 84.1 RIKEN cDNA 2310040G17 gene 1417619_at 4 3.38875643 1.4 2 2.69163229 8.84279E-06 0.0001904 BC056474 15 12.1 Mus musculus cDNA clone 1424117_at H3030A06 3.95752801 2.42838452 1.9 2.2 2.62132809 1.3344E-08 5.66E-07 MGC:67360 IMAGE:6823629, complete cds NA 4 153 guanine nucleotide binding protein, 1454696_at -3.46081884 -4 -1.3 -1.6 -2.6026947 8.58458E-05 0.0012617 beta 1 Gnb1 4 153 guanine nucleotide binding protein, 1417432_a_at H3094D02 -3.13334396 -4 -1.6 -1.7 -2.5946297 1.04542E-05 0.0002202 beta 1 Gadd45gip1 8 84.1 RAD23a homolog (S.
    [Show full text]
  • Anti-GJA4 / Connexin 37 Antibody (ARG58815)
    Product datasheet [email protected] ARG58815 Package: 50 μg anti-GJA4 / Connexin 37 antibody Store at: -20°C Summary Product Description Rabbit Polyclonal antibody recognizes GJA4 / Connexin 37 Tested Reactivity Hu, Ms, Rat Predict Reactivity Hm Tested Application ICC, IHC-Fr, WB Host Rabbit Clonality Polyclonal Isotype IgG Target Name GJA4 / Connexin 37 Species Human Immunogen Synthetic peptide corresponding to aa. 3-17 of Human Connexin 37 (DWGFLEKLLDQVQEH). Conjugation Un-conjugated Alternate Names Connexin-37; Gap junction alpha-4 protein; CX37; Cx37 Application Instructions Application table Application Dilution ICC 0.5 - 1 µg/ml IHC-Fr 1:200 - 1:1000 WB 0.1 - 0.5 µg/ml Application Note * The dilutions indicate recommended starting dilutions and the optimal dilutions or concentrations should be determined by the scientist. Properties Form Liquid Purification Affinity purification with immunogen. Buffer 0.9% NaCl, 0.2% Na2HPO4, 0.05% Thimerosal, 0.05% Sodium azide and 5% BSA. Preservative 0.05% Thimerosal and 0.05% Sodium azide Stabilizer 5% BSA Concentration 0.5 mg/ml Storage instruction For continuous use, store undiluted antibody at 2-8°C for up to a week. For long-term storage, aliquot and store at -20°C or below. Storage in frost free freezers is not recommended. Avoid repeated freeze/thaw cycles. Suggest spin the vial prior to opening. The antibody solution should be gently mixed before use. www.arigobio.com 1/3 Note For laboratory research only, not for drug, diagnostic or other use. Bioinformation Gene Symbol GJA4 Gene Full Name gap junction protein, alpha 4, 37kDa Background This gene encodes a member of the connexin gene family.
    [Show full text]