The Role of Sorting Nexins in Antigen Presentation
|
|
- Kelly Robertson
- 6 years ago
- Views:
Transcription
1 The Role of Sorting Nexins in Antigen Presentation Chng X.R.J 1 and Wong S.H. 2 Department of Microbiology Yong Loo Lin School of Medicine, National University of Singapore Block MD4, 5 Science Drive 2, Singapore ABSTRACT The Sorting Nexins (SNX) are a family of proteins that possess a phoxhomology (PX) domain and which may possess sorting functions. It has been shown in recent studies that some SNX have a role to play in the retromer, a complex that rescues receptors from destruction in the lysosomes and transports these receptors to the golgi. This paper postulates that there may be a link between SNX 5 and 6 and antigen presentation, particularly in the degradation of antigens. However, due to time constraints, this paper only presents the basic groundwork that was laid that can act as a platform to test this postulate in the near future. INTRODUCTION The Sorting Nexin (SNX) Family is a group of cytoplasmic, membraneassociated proteins that are involved in endocytosis and protein trafficking in the cell 1,2,. These SNXs possess a phox-homology (PX) domain that binds to phosphotidylinositol phosphates 1,2,3. More recently, studies have shown that some SNX may be involved with the retromer, a complex that recycles receptors from the endosomes to the golgi, effectively rescuing these receptors from destruction in the lysosomes. The yeast retromer is well studied and consists of Vps35, Vps26, Vps29p and a Vps5p-Vps17 dimer. The former three components have a role in structural identification, where as 1 Student 2 Supervisor
2 the dimer has a structural role in membrane deformation. It was subsequently found that SNX 1 is the mammalian ortholog of Vps5p 2,3,4,5. It was also recently postulated that SNX 5 and SNX 6 may be involved with the retromer. When tested, it was found that repressing SNX 5 and 6 resulted in a phenotype similar to that of repressing known retromer constituents. Also, it was noted that SNX 5 and 6 could co-localise with SNX 1; and more remarkably, the repression of SNX 5 and 6 induced a loss of SNX 1. The paper concluded that SNX 5 and 6 might be an ortholog for Vps17 5. SNX 1 is a relatively well-studied member of the SNX family and it was discovered that it has a role in antigen presentation. It was initially found that Shiga toxin was partially transported to the golgi via SNX 1-labelled endosomes, and that when SNX 1 was suppressed in the cell, Shiga toxin was not as effectively transported. However, the transport of Shiga toxin was not exclusive to SNX 1- labelled endosomes 7. As such, since SNX 1 has been demonstrated to be part of the retromer, and since SNX 5 and 6 have probable roles in the retromer, there might be a likely link between SNX 5 and 6 and antigen presentation. As the retromer is involved in endosome-to-golgi transport, there is a possibility that SNX 5 and 6 may be involved in the trafficking of antigens to the lysosome for degradation during antigen processing, an essential part of antigen presentation. One way of assaying the impact of SNX 5 and 6 on antigen degradation is to monitor the effect SNX 5 and SNX 6 over-expression has on the distribution and degradation of scavenger receptors such as CD204 in macrophages. CD204 is of interest as it is a receptor specific for the uptake of Escherichia coli by macrophages. The qualitative and quantitative assays involving CD204 internalisation were set up, and the endogenous expression of SNX 5 and 6 in RAW cells was studied and attempts were made to overexpress them. MATERIALS AND METHODS The methods used in this study were western blot and immunofluorescence. Western blot was employed to quantitate the internalisation of CD204, while
3 immunofluorescence was used to visualise the internalisation of CD204 and depict endogenous SNX 5 and 6 expression. Transfection methods were used to attempt to increase overexpress the SNX 5 and 6 levels within RAW cells. Immunofluorescence Rat anti-mouse CD204 antibodies from AbD Serotec and goat anti-mouse antibodies for SNX 5 and 6 were used in these assays. The RAW cells were fixed with 4% paraformaldehyde and permeabilised with 0.1% Saponin or 0.1% Triton-X. Following this, RAW cells were incubated in primary antibody and subsequently in fluorescent-labelled secondary antibody. The slides were mounted in medium containing DAPI and subsequently viewed. Transfection to overexpress SNX 5 and 6 Transfection was carried out using Lipofectamine 2000 TM and Effectene TM with differing concentrations of DNA (Lipofectamine 2000 TM : 4ug and 8ug of DNA; Effectene TM : 0.4ug, 0.8ug, 1.6ug, 3.2ug, 4.8ug, 6.4ug) in RAW cells. Effectene TM was purchased from QIAGEN, and Lipofectamine 2000 TM from Invitrogen. Western blotting The antibody that was used in this assay was a rat anti-mouse CD204 antibody from AbD Serotec. RESULTS AND DISCUSSION An assay system to qualify and quantify the internalisation of CD204 was constructed. As we expect the overexpression of SNX 5 and 6 to have an impact on the internalisation of CD204, this platform is essential to assessing the effects the overexpression of SNX 5 and 6 on CD204 internalisation. SNX 5 and 6 were observed and efforts were made to over express them; however, the latter step proved problematic and unsuccessful. Quantitative analysis of CD204 internalisation This kinetics experiment was conducted using western blot at time points of 0, 5, 10, 15, 30 and 60 minutes on RAW cells with an anti-cd204 antibody. It was
4 observed that there was rapid degradation of CD204, with discernable difference being noted five minutes after the commencement of the experiment. This assay can be used on RAW cells overexpressing SNX 5 and 6 to determine the effect, if any, that SNX 5 and 6 have on the degradation of CD204. Qualitative analysis of CD204 internalisation A kinetics experiment involving two time points (0 min and 60 min) was carried out to observe the internalisation of CD204 and an attempt to co-localise CD204 with VAMP 3, and early endosome marker was also done. It was observed that CD204 on the surface was transported into endosomes after an hour, with no colocalisation with VAMP 3 i.e. CD204 does not reside in early endosomes an hour after internalisation. More investigations can be done on this experiment, for example, co-localisation with other markers e.g. VAMP 8, and it could be repeated at more time points to more clearly see the internalisation. Endogenous expression of SNX 5 and 6 SNX 5 and 6 were observed, and co-localisation of SNX 6 with vti1a was carried out. SNX 5 was not co-localised with vti1a as the antibodies available were not compatible. Vti1a is a marker for endosomes and the golgi 6,7. It was noted that there is co-localisation between SNX 6 and vti1a, and it seems that both SNX 5 and 6 are predominantly expressed in the endosomes, reflecting what was previously reported in the literature. Transfection of SNX 5 and 6 SNX 5 and 6 were overexpressed transiently in RAW cells by transfection with plasmid DNA. However, the transfection efficiencies remained low, despite changing various parameters like the reagents used, the amount of DNA added and the purity of the DNA. The experiment was unable to proceed from there, but to circumvent this problem, a stable clone that overexpresses SNX 5 or 6 can be selected and obtained. REFERENCES
5 1. Worby CA, Dixon JE. Sorting out the cellular functions of sorting nexins. Nat Rev Mol Cell Biol Dec;3(12): Carlton JG, Cullen PJ. Sorting nexins. Curr Biol Oct 25;15(20):R Carlton J, Bujny M, Rutherford A, Cullen P. Sorting nexins--unifying trends and new perspectives. Traffic Feb;6(2): Seaman MN. Recycle your receptors with retromer. Trends Cell Biol Feb;15(2): Verges M. Retromer and sorting nexins in development. Front Biosci May 1;12: Kreykenbohm V.; Wenzel D.; Antonin W.; Atlachkine V.; Fischer von Mollard G. The SNAREs vti1a and vti1b have distinct localization and SNARE complex partners. European Journal of Cell Biology, Volume 81, Number 5, May 2002, pp (8) 7. Wolfram Antonin, Dietmar Riedel, Gabriele Fischer von Mollard. The SNARE Vti1a-beta Is Localized to Small Synaptic Vesicles and Participates in a Novel SNARE Complex. The Journal of Neuroscience, August 1, 2000, 20(15):
Rab9-dependent retrograde transport and endosomal sorting of the endopeptidase furin
Research Article 2401 Rab9-dependent retrograde transport and endosomal sorting of the endopeptidase furin Pei Zhi Cheryl Chia, Isabelle Gasnereau, Zi Zhao Lieu and Paul A. Gleeson* The Department of Biochemistry
More informationLysosomes and endocytic pathways 9/27/2012 Phyllis Hanson
Lysosomes and endocytic pathways 9/27/2012 Phyllis Hanson General principles Properties of lysosomes Delivery of enzymes to lysosomes Endocytic uptake clathrin, others Endocytic pathways recycling vs.
More informationIntracellular Vesicular Traffic Chapter 13, Alberts et al.
Intracellular Vesicular Traffic Chapter 13, Alberts et al. The endocytic and biosynthetic-secretory pathways The intracellular compartments of the eucaryotic ell involved in the biosynthetic-secretory
More informationgenome edited transient transfection, CMV promoter
Supplementary Figure 1. In the absence of new protein translation, overexpressed caveolin-1-gfp is degraded faster than caveolin-1-gfp expressed from the endogenous caveolin 1 locus % loss of total caveolin-1-gfp
More informationIP: anti-gfp VPS29-GFP. IP: anti-vps26. IP: anti-gfp - + +
FAM21 Strump. WASH1 IP: anti- 1 2 3 4 5 6 FAM21 Strump. FKBP IP: anti-gfp VPS29- GFP GFP-FAM21 tail H H/P P H H/P P c FAM21 FKBP Strump. VPS29-GFP IP: anti-gfp 1 2 3 FKBP VPS VPS VPS VPS29 1 = VPS29-GFP
More informationAn analysis of sub-cellular localization of host sorting nexin 4 and 7 in Coxiella burnetii infected Hela cells
An analysis of sub-cellular localization of host sorting nexin 4 and 7 in Coxiella burnetii infected Hela cells Authors: Sydni Smith, Saugata Mahapatra PhD., Edward Shaw PhD.* Department of Microbiology
More informationTHE ROLE OF ALTERED CALCIUM AND mtor SIGNALING IN THE PATHOGENESIS OF CYSTINOSIS
Research Foundation, 18 month progress report THE ROLE OF ALTERED CALCIUM AND mtor SIGNALING IN THE PATHOGENESIS OF CYSTINOSIS Ekaterina Ivanova, doctoral student Elena Levtchenko, MD, PhD, PI Antonella
More informationMany G protein-coupled receptors (GPCRs) 2 are rapidly endocytosed after agonist binding, but the pathway of postendocytic
THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 282, NO. 40, pp. 29646 29657, October 5, 2007 2007 by The American Society for Biochemistry and Molecular Biology, Inc. Printed in the U.S.A. Hepatocyte Growth
More information1) INVESTIGATION OF THE CLATHRIN-DEPENDENT INTERNALISATION OF CYTOTOXIC T LYMPHOCYTE ANTIGEN-4
1) INVESTIGATION OF THE CLATHRIN-DEPENDENT INTERNALISATION OF CYTOTOXIC T LYMPHOCYTE ANTIGEN-4 2) ANALYSIS OF THE ANTIGEN-SPECIFIC CYTOKINE RESPONSES OF CD4 + T-LYMPHOCYTES This thesis is submitted by
More information04_polarity. The formation of synaptic vesicles
Brefeldin prevents assembly of the coats required for budding Nocodazole disrupts microtubules Constitutive: coatomer-coated Selected: clathrin-coated The formation of synaptic vesicles Nerve cells (and
More informationStudying dynamic protein:protein interactions in vesicle trafficking. David Owen CIMR University of Cambridge
Studying dynamic protein:protein interactions in vesicle trafficking David Owen CIMR University of Cambridge The importance of vesicle trafficking in mammalian cells Transducing signals small molecules/nutrients
More informationUNIVERSITY OF YORK BSc Stage 2 Degree Examinations Department: BIOLOGY. Title of Exam: Cell Biology
Examination Candidate Number: Desk Number: UNIVERSITY OF YORK BSc Stage 2 Degree Examinations 2017-18 Department: BIOLOGY Title of Exam: Cell Biology Time allowed: 1 hour and 30 minutes Total marks available
More informationFig. S1. Subcellular localization of overexpressed LPP3wt-GFP in COS-7 and HeLa cells. Cos7 (top) and HeLa (bottom) cells expressing for 24 h human
Fig. S1. Subcellular localization of overexpressed LPP3wt-GFP in COS-7 and HeLa cells. Cos7 (top) and HeLa (bottom) cells expressing for 24 h human LPP3wt-GFP, fixed and stained for GM130 (A) or Golgi97
More informationChapter 13: Vesicular Traffic
Chapter 13: Vesicular Traffic Know the terminology: ER, Golgi, vesicle, clathrin, COP-I, COP-II, BiP, glycosylation, KDEL, microtubule, SNAREs, dynamin, mannose-6-phosphate, M6P receptor, endocytosis,
More informationFigure S1. Western blot analysis of clathrin RNA interference in human DCs Human immature DCs were transfected with 100 nm Clathrin SMARTpool or
Figure S1. Western blot analysis of clathrin RNA interference in human DCs Human immature DCs were transfected with 100 nm Clathrin SMARTpool or control nontargeting sirnas. At 90 hr after transfection,
More informationCOURSE: Medical Microbiology, MBIM 650/720 - Fall TOPIC: Antigen Processing, MHC Restriction, & Role of Thymus Lecture 12
COURSE: Medical Microbiology, MBIM 650/720 - Fall 2008 TOPIC: Antigen Processing, MHC Restriction, & Role of Thymus Lecture 12 FACULTY: Dr. Mayer Office: Bldg. #1, Rm B32 Phone: 733-3281 Email: MAYER@MED.SC.EDU
More informationOctober 26, Lecture Readings. Vesicular Trafficking, Secretory Pathway, HIV Assembly and Exit from Cell
October 26, 2006 Vesicular Trafficking, Secretory Pathway, HIV Assembly and Exit from Cell 1. Secretory pathway a. Formation of coated vesicles b. SNAREs and vesicle targeting 2. Membrane fusion a. SNAREs
More informationSupplementary Figure 1.TRIM33 binds β-catenin in the nucleus. a & b, Co-IP of endogenous TRIM33 with β-catenin in HT-29 cells (a) and HEK 293T cells
Supplementary Figure 1.TRIM33 binds β-catenin in the nucleus. a & b, Co-IP of endogenous TRIM33 with β-catenin in HT-29 cells (a) and HEK 293T cells (b). TRIM33 was immunoprecipitated, and the amount of
More informationLocalization and Retention of Glycosyltransferases And the Role of Vesicle Trafficking in Glycosylation
Localization and Retention of Glycosyltransferases And the Role of Vesicle Trafficking in Glycosylation Richard Steet, Ph.D. 2/21/17 glycosylation is a non-template derived phenomenon - the presence of
More informationMechanism of Vesicular Transport
Mechanism of Vesicular Transport Transport vesicles play a central role in the traffic of molecules between different membrane-enclosed enclosed compartments. The selectivity of such transport is therefore
More informationThe functional investigation of the interaction between TATA-associated factor 3 (TAF3) and p53 protein
THESIS BOOK The functional investigation of the interaction between TATA-associated factor 3 (TAF3) and p53 protein Orsolya Buzás-Bereczki Supervisors: Dr. Éva Bálint Dr. Imre Miklós Boros University of
More informationThe Molecular Mechanism of Intracellular Membrane Fusion. Richard H. Scheller
The Molecular Mechanism of Intracellular Membrane Fusion Richard H. Scheller The human brain contains approximately 10 15 connections between nerve cells (Figure 2). The specific formation and modulation
More informationProtein Trafficking in the Secretory and Endocytic Pathways
Protein Trafficking in the Secretory and Endocytic Pathways The compartmentalization of eukaryotic cells has considerable functional advantages for the cell, but requires elaborate mechanisms to ensure
More informationLocalization and Retention of Glycosyltransferases And the Role of Vesicle Trafficking in Glycosylation
Localization and Retention of Glycosyltransferases And the Role of Vesicle Trafficking in Glycosylation Richard Steet, Ph.D. 3/8/2011 glycosylation is a non-template derived phenomenon - the presence of
More informationModule 3 Lecture 7 Endocytosis and Exocytosis
Module 3 Lecture 7 Endocytosis and Exocytosis Endocytosis: Endocytosis is the process by which cells absorb larger molecules and particles from the surrounding by engulfing them. It is used by most of
More informationWT MSD MPS-IIIA. Lysosomal ph
7 6 Lysosomal ph 5 4 3 2 1 Supplementary figure S1. Lysosomal ph was measure in, and MPSIIIA MEFs using a Lysosensor yellow/blue-dextran (Molecular Probes) as a lysosomal ph indicator (see methods). P
More informationMultidimensional Tracking of GPCR Signaling with Proximity- Labeling. Technical Journal Club Anna Henzi,
Multidimensional Tracking of GPCR Signaling with Proximity- Labeling Technical Journal Club Anna Henzi, 11.07.2017 Proximity-dependent labeling methods Enzymes create radicals Proximity-dependent biotin
More informationVesicle Transport. Vesicle pathway: many compartments, interconnected by trafficking routes 3/17/14
Vesicle Transport Vesicle Formation Curvature (Self Assembly of Coat complex) Sorting (Sorting Complex formation) Regulation (Sar1/Arf1 GTPases) Fission () Membrane Fusion SNARE combinations Tethers Regulation
More informationChapter 3. Expression of α5-megfp in Mouse Cortical Neurons. on the β subunit. Signal sequences in the M3-M4 loop of β nachrs bind protein factors to
22 Chapter 3 Expression of α5-megfp in Mouse Cortical Neurons Subcellular localization of the neuronal nachr subtypes α4β2 and α4β4 depends on the β subunit. Signal sequences in the M3-M4 loop of β nachrs
More informationExtracellular vesicle budding is inhibited by redundant regulators of TAT-5 flippase localization and phospholipid asymmetry
Extracellular vesicle budding is inhibited by redundant regulators of TAT-5 flippase localization and phospholipid asymmetry Katharina. eer a, Jennifer Rivas-Castillo a, Kenneth Kuhn a, Gholamreza Fazeli
More informationSupplemental information
Carcinoemryonic antigen-related cell adhesion molecule 6 (CEACAM6) promotes EGF receptor signaling of oral squamous cell carcinoma metastasis via the complex N-glycosylation y Chiang et al. Supplemental
More informationIntracellular Vesicle Trafficking
Intracellular Vesicle Trafficking Chi-Kuang Yao (IBC, Academia Sinica) 11-6-2017 ckyao@gate.sinica.edu.tw 1 Compartmentalization makes difference between bacteria and yeast 1. More compartments with specific
More informationSupplementary Information
Supplementary Information Supplementary Figure 1: cholesterol manipulation alters the positioning of autophagosomes in cells, related to figure 1. (a) HeLa cells were treated for 24h under conditions reducing
More informationSUPPLEMENTARY INFORMATION
Figure S1 Treatment with both Sema6D and Plexin-A1 sirnas induces the phenotype essentially identical to that induced by treatment with Sema6D sirna alone or Plexin-A1 sirna alone. (a,b) The cardiac tube
More informationSupporting Information. FADD regulates NF-кB activation and promotes ubiquitination of cflip L to induce. apoptosis
1 2 Supporting Information 3 4 5 FADD regulates NF-кB activation and promotes ubiquitination of cflip L to induce apoptosis 6 7 Kishu Ranjan and Chandramani Pathak* 8 9 Department of Cell Biology, School
More informationAntigen presenting cells
Antigen recognition by T and B cells - T and B cells exhibit fundamental differences in antigen recognition - B cells recognize antigen free in solution (native antigen). - T cells recognize antigen after
More informationMicroRNA sponges: competitive inhibitors of small RNAs in mammalian cells
MicroRNA sponges: competitive inhibitors of small RNAs in mammalian cells Margaret S Ebert, Joel R Neilson & Phillip A Sharp Supplementary figures and text: Supplementary Figure 1. Effect of sponges on
More informationSupporting Information
Supporting Information Fig. S1. Overexpression of Rpr causes progenitor cell death. (A) TUNEL assay of control intestines. No progenitor cell death could be observed, except that some ECs are undergoing
More informationReady-to-use Lentiviral Particles for intracelular labeling
Ready-to-use Lentiviral Particles for intracelular labeling (LocLight TM Living cell imaging lentivirus for sub-cellular localization) LocLight TM cell organelle labeling lentivirus are provided as 200ul/per
More informationT H E J O U R N A L O F C E L L B I O L O G Y
T H E J O U R N A L O F C E L L B I O L O G Y Supplemental material Krenn et al., http://www.jcb.org/cgi/content/full/jcb.201110013/dc1 Figure S1. Levels of expressed proteins and demonstration that C-terminal
More informationSUPPLEMENTARY FIGURES
SUPPLEMENTARY FIGURES Supplementary Figure 1. (A) Left, western blot analysis of ISGylated proteins in Jurkat T cells treated with 1000U ml -1 IFN for 16h (IFN) or left untreated (CONT); right, western
More informationEndocytosis and retrograde transport of Shiga toxin and ricin
Audrun Utskarpen Endocytosis and retrograde transport of Shiga toxin and ricin Centre for Cancer Biomedicine, University of Oslo Department of Biochemistry, Institute for Cancer Research, Norwegian Radium
More informationThe Major Histocompatibility Complex (MHC)
The Major Histocompatibility Complex (MHC) An introduction to adaptive immune system before we discuss MHC B cells The main cells of adaptive immune system are: -B cells -T cells B cells: Recognize antigens
More informationT H E J O U R N A L O F C E L L B I O L O G Y
Supplemental material Chairoungdua et al., http://www.jcb.org/cgi/content/full/jcb.201002049/dc1 T H E J O U R N A L O F C E L L B I O L O G Y Figure S1. Expression of CD9 and CD82 inhibits Wnt/ -catenin
More informationLipidne mikrodomene. funkcija
Lipidne mikrodomene funkcija 1 Cellular processes involving lipid rafts - Signal transduction - Protein and lipid trafficking and sorting - Endosome(clathrin)-independent endocytosis: - potocytosis and
More informationFOXO Reporter Kit PI3K/AKT Pathway Cat. #60643
Data Sheet FOXO Reporter Kit PI3K/AKT Pathway Cat. #60643 Background The PI3K/AKT signaling pathway is essential for cell growth and survival. Disruption of this pathway or its regulation has been linked
More informationLecture Readings. Vesicular Trafficking, Secretory Pathway, HIV Assembly and Exit from Cell
October 26, 2006 1 Vesicular Trafficking, Secretory Pathway, HIV Assembly and Exit from Cell 1. Secretory pathway a. Formation of coated vesicles b. SNAREs and vesicle targeting 2. Membrane fusion a. SNAREs
More informationProject report October 2012 March 2013 CRF fellow: Principal Investigator: Project title:
Project report October 2012 March 2013 CRF fellow: Gennaro Napolitano Principal Investigator: Sergio Daniel Catz Project title: Small molecule regulators of vesicular trafficking to enhance lysosomal exocytosis
More information7.06 Cell Biology EXAM #3 April 24, 2003
7.06 Spring 2003 Exam 3 Name 1 of 8 7.06 Cell Biology EXAM #3 April 24, 2003 This is an open book exam, and you are allowed access to books and notes. Please write your answers to the questions in the
More informationSupplementary Figure 1. Spatial distribution of LRP5 and β-catenin in intact cardiomyocytes. (a) and (b) Immunofluorescence staining of endogenous
Supplementary Figure 1. Spatial distribution of LRP5 and β-catenin in intact cardiomyocytes. (a) and (b) Immunofluorescence staining of endogenous LRP5 in intact adult mouse ventricular myocytes (AMVMs)
More informationChapter 1: Vesicular traffic. Biochimica cellulare parte B 2017/18
Chapter 1: Vesicular traffic Biochimica cellulare parte B 2017/18 Major Protein-sorting pathways in eukaryotic cells Secretory and endocytic pathways Unifying principle governs all protein trafficking
More informationGeneral information. Cell mediated immunity. 455 LSA, Tuesday 11 to noon. Anytime after class.
General information Cell mediated immunity 455 LSA, Tuesday 11 to noon Anytime after class T-cell precursors Thymus Naive T-cells (CD8 or CD4) email: lcoscoy@berkeley.edu edu Use MCB150 as subject line
More informationHEK293FT cells were transiently transfected with reporters, N3-ICD construct and
Supplementary Information Luciferase reporter assay HEK293FT cells were transiently transfected with reporters, N3-ICD construct and increased amounts of wild type or kinase inactive EGFR. Transfections
More informationSupplemental Materials. STK16 regulates actin dynamics to control Golgi organization and cell cycle
Supplemental Materials STK16 regulates actin dynamics to control Golgi organization and cell cycle Juanjuan Liu 1,2,3, Xingxing Yang 1,3, Binhua Li 1, Junjun Wang 1,2, Wenchao Wang 1, Jing Liu 1, Qingsong
More informationMISSION: understanding the mechanisms of therapeutic strategies
Telethon Institute of Genetics and Medicine MISSION: understanding the mechanisms of genetic diseases to develop preventive and therapeutic strategies G E N O T Y P E G E Researcher N 1 3 5 O T Y P E 8
More informationThe regulation and function of Class III PI3Ks: novel roles for Vps34
Biochem. J. (2008) 410, 1 17 (Printed in Great Britain) doi:10.1042/bj20071427 1 REVIEW ARTICLE The regulation and function of Class III PI3Ks: novel roles for Vps34 Jonathan M. BACKER 1 Department of
More informationLab module 6a Receptor-mediated endocytosis
Goal for the module Lab module 6a Receptor-mediated endocytosis To follow cell surface receptors as they are internalized. Pre-lab homework Read about receptor-mediated endocytosis and other forms of internalization
More information7.06 Spring of PROBLEM SET #6
7.6 Spring 23 1 of 6 7.6 PROBLEM SET #6 1. You are studying a mouse model of hypercholesterolemia, a disease characterized by high levels of cholesterol in the blood. In normal cells, LDL particles in
More informationFigure S1. Sorting nexin 9 (SNX9) specifically binds psmad3 and not psmad 1/5/8. Lysates from AKR-2B cells untreated (-) or stimulated (+) for 45 min
Figure S1. Sorting nexin 9 (SNX9) specifically binds psmad3 and not psmad 1/5/8. Lysates from AKR2B cells untreated () or stimulated () for 45 min with 5 ng/ml TGFβ or 10 ng/ml BMP4 were incubated with
More informationLong term stability of isolated human serum derived exosomes
Long term stability of isolated human serum derived exosomes Candice de Boer (PhD student) Regenerative Medicine Laboratory Supervisor: Associate Professor Neil Davies Exosomes First discovered during
More informationSignificance of the MHC
CHAPTER 8 Major Histocompatibility Complex (MHC) What is is MHC? HLA H-2 Minor histocompatibility antigens Peter Gorer & George Sneell (1940) Significance of the MHC role in immune response role in organ
More informationLDLR-related protein 10 (LRP10) regulates amyloid precursor protein (APP) trafficking and processing: evidence for a role in Alzheimer s disease
Brodeur et al. Molecular Neurodegeneration 2012, 7:31 RESEARCH ARTICLE Open Access LDLR-related protein 10 (LRP10) regulates amyloid precursor protein (APP) trafficking and processing: evidence for a role
More informationAppendix: Subcellular localization of MIG-14::GFP in C. elegans body wall muscle cells. Pei-Tzu Yang and Hendrik C. Korswagen
Appendix: Subcellular localization of MIG-14::GFP in C. elegans body wall muscle cells Pei-Tzu Yang and Hendrik C. Korswagen 129 SUBCELLULAR LOCALIZATION OF MIG-14 Abstract MIG-14/Wls is a Wnt binding
More informationSupplementary Figure 1. PAQR3 knockdown inhibits SREBP-2 processing in CHO-7 cells CHO-7 cells were transfected with control sirna or a sirna
Supplementary Figure 1. PAQR3 knockdown inhibits SREBP-2 processing in CHO-7 cells CHO-7 cells were transfected with control sirna or a sirna targeted for hamster PAQR3. At 24 h after the transfection,
More informationThe role of the scaffolding protein Tks5 in EGF signaling
The role of the scaffolding protein Tks5 in EGF signaling PhD Thesis Anna Fekete Doctoral School in Biology Head of the School: Dr. Anna Erdei Structural Biochemistry Doctoral Program Head of the Program:
More informationPhosphoinositide Signaling
Phosphoinositide Signaling Pietro De Camilli Department of Cell Biology Howard Hughes Medical Institute Program in Cellular Neuroscience, Neurodegeneration and Repair Yale University School of Medicine
More informationSupplementary Figure S1. Venn diagram analysis of mrna microarray data and mirna target analysis. (a) Western blot analysis of T lymphoblasts (CLS)
Supplementary Figure S1. Venn diagram analysis of mrna microarray data and mirna target analysis. (a) Western blot analysis of T lymphoblasts (CLS) and their exosomes (EXO) in resting (REST) and activated
More informationHuman SH-SY5Y neuroblastoma cells (A.T.C.C., Manassas, VA) were cultured in DMEM, F-12
SUPPLEMENTARY METHODS Cell cultures Human SH-SY5Y neuroblastoma cells (A.T.C.C., Manassas, VA) were cultured in DMEM, F-12 Ham with 25 mm HEPES and NaHCO 3 (1:1) and supplemented with 10% (v/v) FBS, 1.0
More informationSorting nexin 4 and amphiphysin 2, a new partnership between endocytosis and intracellular trafficking
Research Article 1937 Sorting nexin 4 and amphiphysin 2, a new partnership between endocytosis and intracellular trafficking Corinne Leprince 1, *, Erwan Le Scolan 1, Brigitte Meunier 1, Vincent Fraisier
More informationEndocytosis and Intracellular Trafficking of Notch and Its Ligands
CHA P T E R F IVE Endocytosis and Intracellular Trafficking of Notch and Its Ligands Shinya Yamamoto, *,1 Wu-Lin Charng, *,1 and Hugo J. Bellen *,,, Contents 1. Notch Signaling and its Regulation by Endocytosis
More informationSestrin2 and BNIP3 (Bcl-2/adenovirus E1B 19kDa-interacting. protein3) regulate autophagy and mitophagy in renal tubular cells in. acute kidney injury
Sestrin2 and BNIP3 (Bcl-2/adenovirus E1B 19kDa-interacting protein3) regulate autophagy and mitophagy in renal tubular cells in acute kidney injury by Masayuki Ishihara 1, Madoka Urushido 2, Kazu Hamada
More informationSUPPLEMENTARY INFORMATION. Supp. Fig. 1. Autoimmunity. Tolerance APC APC. T cell. T cell. doi: /nature06253 ICOS ICOS TCR CD28 TCR CD28
Supp. Fig. 1 a APC b APC ICOS ICOS TCR CD28 mir P TCR CD28 P T cell Tolerance Roquin WT SG Icos mrna T cell Autoimmunity Roquin M199R SG Icos mrna www.nature.com/nature 1 Supp. Fig. 2 CD4 + CD44 low CD4
More informationReviewers' comments: Reviewer #1 Expert in EAE and IL-17a (Remarks to the Author):
Reviewers' comments: Reviewer #1 Expert in EAE and IL-17a (Remarks to the Author): This study shows that the inducible camp early repressor (ICER) is involved in development of Th17 cells that are pathogenic
More informationHLA and antigen presentation. Department of Immunology Charles University, 2nd Medical School University Hospital Motol
HLA and antigen presentation Department of Immunology Charles University, 2nd Medical School University Hospital Motol MHC in adaptive immunity Characteristics Specificity Innate For structures shared
More informationLipids and Membranes
Lipids and Membranes Presented by Dr. Mohammad Saadeh The requirements for the Pharmaceutical Biochemistry I Philadelphia University Faculty of pharmacy Membrane transport D. Endocytosis and Exocytosis
More informationFang et al. NMuMG. PyVmT unstained Anti-CCR2-PE MDA-MB MCF MCF10A
A NMuMG PyVmT 16.5+.5 47.+7.2 Fang et al. unstained Anti-CCR2-PE 4T1 Control 37.6+6.3 56.1+.65 MCF1A 16.1+3. MCF-7 3.1+5.4 MDA-M-231 42.1+5.5 unstained Secondary antibody only Anti-CCR2 SUPPLEMENTAL FIGURE
More informationPROTEIN TRAFFICKING. Dr. SARRAY Sameh, Ph.D
PROTEIN TRAFFICKING Dr. SARRAY Sameh, Ph.D Overview Proteins are synthesized either on free ribosomes or on ribosomes bound to endoplasmic reticulum (RER). The synthesis of nuclear, mitochondrial and peroxisomal
More informationOxford Expression Technologies Ltd
Oxford Expression Technologies Ltd Founded in 2007 as a spin out from Oxford Brookes University and Natural Environment Research Council Technology based on the insect baculovirus expression vectors (BEVs)
More informationSupplementary information. MARCH8 inhibits HIV-1 infection by reducing virion incorporation of envelope glycoproteins
Supplementary information inhibits HIV-1 infection by reducing virion incorporation of envelope glycoproteins Takuya Tada, Yanzhao Zhang, Takayoshi Koyama, Minoru Tobiume, Yasuko Tsunetsugu-Yokota, Shoji
More informationTITLE: Effect of MUC1 Expression on EGFR Endocytosis and Degradation in Human Breast Cancer Cell Lines
AD AWARD NUMBER: W81XWH-06-1-0464 TITLE: Effect of MUC1 Expression on EGFR Endocytosis and Degradation in Human Breast Cancer Cell Lines PRINCIPAL INVESTIGATOR: Rachid M El Bejjani CONTRACTING ORGANIZATION:
More informationCellular Mechanisms for the Release of Exosomes Containing the Amyloid Precursor Protein
Cellular Mechanisms for the Release of Exosomes Containing the Amyloid Precursor Protein Master s Thesis Presented to The Faculty of the Graduate School of Arts and Sciences Brandeis University Interdepartmental
More informationThe IL-7 Receptor A Key Factor in HIV Pathogenesis
The IL-7 Receptor A Key Factor in HIV Pathogenesis Paul MacPherson PhD, MD, FRCPC Associate Professor Division of Infectious Diseases Ottawa Hospital, General Campus Ottawa Hospital Research Institute
More informationGLYCATION OF PROTEINS IN ESCHERICHIA COLI: EFFECT OF NUTRIENT BROTH INGREDIENTS ON GLYCATION
Industry GLYCATION OF PROTEINS IN ESCHERICHIA COLI: EFFECT OF NUTRIENT BROTH INGREDIENTS ON GLYCATION R. Dimitrova, R. Mironova, I. Ivanov Institute of Molecular biology, Bulgarian Academy of Sciences
More informationeffects on organ development. a-f, Eye and wing discs with clones of ε j2b10 show no
Supplementary Figure 1. Loss of function clones of 14-3-3 or 14-3-3 show no significant effects on organ development. a-f, Eye and wing discs with clones of 14-3-3ε j2b10 show no obvious defects in Elav
More informationThe Role of the Scaffolding Protein Tks4 in EGF Signaling. Gábor Bőgel. Semmelweis University Molceular Medicine PhD School
The Role of the Scaffolding Protein Tks4 in EGF Signaling PhD Theses Gábor Bőgel Semmelweis University Molceular Medicine PhD School Supervisor: Dr. László Buday MD, D.Sc. Official reviewers: Dr. Beáta
More information(a) Significant biological processes (upper panel) and disease biomarkers (lower panel)
Supplementary Figure 1. Functional enrichment analyses of secretomic proteins. (a) Significant biological processes (upper panel) and disease biomarkers (lower panel) 2 involved by hrab37-mediated secretory
More information2013 John Wiley & Sons, Inc. All rights reserved. PROTEIN SORTING. Lecture 10 BIOL 266/ Biology Department Concordia University. Dr. S.
PROTEIN SORTING Lecture 10 BIOL 266/4 2014-15 Dr. S. Azam Biology Department Concordia University Introduction Membranes divide the cytoplasm of eukaryotic cells into distinct compartments. The endomembrane
More informationSupplementary Figure 1 Expression of Crb3 in mouse sciatic nerve: biochemical analysis (a) Schematic of Crb3 isoforms, ERLI and CLPI, indicating the
Supplementary Figure 1 Expression of Crb3 in mouse sciatic nerve: biochemical analysis (a) Schematic of Crb3 isoforms, ERLI and CLPI, indicating the location of the transmembrane (TM), FRM binding (FB)
More informationsupplementary information
DOI: 10.1038/ncb2133 Figure S1 Actomyosin organisation in human squamous cell carcinoma. (a) Three examples of actomyosin organisation around the edges of squamous cell carcinoma biopsies are shown. Myosin
More information40s 50s. 70s. Membrane Rafts
40s 50s 70s Membrane Rafts Membrane Microdomains Raft is a specific type of microdomain sphingolipid/cholesterol rich region Separation of discrete liquid-ordered and liquid-disordered phase domains occurring
More informationSupplementary Material and Methods
Online Supplement Kockx et al, Secretion of Apolipoprotein E from Macrophages 1 Supplementary Material and Methods Cloning of ApoE-GFP Full-length human apoe3 cdna (pcdna3.1/zeo + -apoe) was kindly provided
More informationSupplemental Material for. Figure S1. Identification of TetR responsive promoters in F. novicida and E. coli.
Supplemental Material for Synthetic promoters functional in Francisella novicida and Escherichia coli Ralph L. McWhinnie and Francis E. Nano Department of Biochemistry and Microbiology, University of Victoria,
More informationOrganization of ATPases
The Primary Active Transporter II: The ATPase Objectives: Organization P type with NPA domains Proton pumps of the rotary V type ATPase 1 Organization of P type, solute transport, found in plasma membranes
More informationSUPPLEMENTARY INFORMATION
doi: 1.138/nature775 4 O.D. (595-655) 3 1 -ζ no antibody isotype ctrl Plated Soluble 1F6 397 7H11 Supplementary Figure 1 Soluble and plated anti- Abs induce -! signalling. B3Z cells stably expressing!
More informationHLA and antigen presentation. Department of Immunology Charles University, 2nd Medical School University Hospital Motol
HLA and antigen presentation Department of Immunology Charles University, 2nd Medical School University Hospital Motol MHC in adaptive immunity Characteristics Specificity Innate For structures shared
More informationTitle of file for HTML: Supplementary Information Description: Supplementary Figures and Supplementary Table
Title of file for HTML: Supplementary Information Description: Supplementary Figures and Supplementary Table Title of file for HTML: Peer Review File Description: Innate Scavenger Receptor-A regulates
More informationHomework Hanson section MCB Course, Fall 2014
Homework Hanson section MCB Course, Fall 2014 (1) Antitrypsin, which inhibits certain proteases, is normally secreted into the bloodstream by liver cells. Antitrypsin is absent from the bloodstream of
More informationReconstitution of GABAergic Postsynapses in Host Cells
University of Connecticut OpenCommons@UConn Honors Scholar Theses Honors Scholar Program Spring 4-27-2018 Reconstitution of GABAergic Postsynapses in Host Cells Karthik Kanamalla karthik.kanamalla@gmail.com
More informationAnti-DC-SIGN/CD209 murine monoclonal antibodies
Anti-DC-SIGN/CD209 murine monoclonal antibodies DC-SIGN (DC Specific, ICAM-3 Grabbing, Nonintegrin) / CD209 and L-SIGN (liver/lymph node-specific ICAM-3-grabbing nonintegrin CD299/ DC-SIGNR (DC-SIGN-related
More informationTSH Receptor Monoclonal Antibody (49) Catalog Number MA3-218 Product data sheet
Website: thermofisher.com Customer Service (US): 1 800 955 6288 ext. 1 Technical Support (US): 1 800 955 6288 ext. 441 TSH Receptor Monoclonal Antibody (49) Catalog Number MA3-218 Product data sheet Details
More information