DOCSLIB.ORG

Supplemental Figure 1: Immunoelectron Microscopy of Ribeye on the Harvested Sucrose Pellet from the Gradient Centrifugation Shown in Multiple Panels

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Supplemental Figure 1: Immunoelectron Microscopy of Ribeye on the Harvested Sucrose Pellet from the Gradient Centrifugation Shown in Multiple Panels Supplemental Figure 1: Immunoelectron microscopy of ribeye on the harvested sucrose pellet from the gradient centrifugation shown in multiple panels. Supplemental Figure 2: Immunoelectron microscopy of VAMP2 on the harvested 200-400 mM sucrose interface sample shown in multiple panels. Supplemental Figure 3: A. Western blots indicate the immunoprecipitation of ribeye and CtBP2 with the CtBP2 antibody. Also coimmunoprecipitation of CtBP1, actin, myosin 9H, spectrin and α-tubulin is shown. B. Western blots indicate the immunoprecipitation of only ribeye with the AD antibody and the coimmunoprecipitation of PRPF39 is indicated. Supplemental Figure 4. Colocalization of ribeye with various presynaptic proteins. The labeling of individual proteins is presented in grayscale and merged panels are depicted in color. Double-labeling with ribeye-specific antiserum reveals the localization with respect to synaptic ribbons of SNARE proteins such as syntaxin 1 (A-C), SNAP25 (D-F), VAMP2 (G-I), syntaxin 6 (J-L), SNAP23 (M-O), VAMP7 (P-R), VAP-33 (S-U); SNARE dissociation molecules such as α-SNAP, β-SNAP (V-X), NSF (Y-A′); hair-cell specific proteins such as glutamate receptors 2 and 3 (GluR2 and GluR3) (B′-D′), glutamate transporter vGluT3 (E′-G′), hair-cell specific Ca2+ channel Cav1.3 (H′-J′), calcium sensor otoferlin (K′-M′); and other synaptic proteins such as rabphilin (N′-P′), munc 18 (Q′-S′), and synapsin (T′-V′). Supplemental Figure 5. Colocalization of ribeye with endocytotic and vesicle- trafficking proteins. The labeling of individual proteins is presented in grayscale and merged panels are depicted in color. Double-labeling with ribeye-specific antiserum reveals the localization of the endocytic proteins clathrin (A-C), dynamin (D-F), SCAMP1 (G-I) and endophilin (J-L) and of the trafficking proteins Sec13 (M-O), β-COP (P-R), VCP (S-U), and GAPDH (V-X) with respect to synaptic ribbons. Supplemental Figure 6. Colocalization of ribeye with cytoskeletal proteins. The labeling of individual proteins is presented in grayscale and merged panels are depicted in color. Double-labeling with ribeye-specific antiserum reveals the localization of the cytoskeletal proteins α-tubulin (A-C), spectrin (D-F), MYH9 (G-I), β-catenin (J-L), NF200 (M-O), and KIF3A (P-R) with respect to synaptic ribbons. Supplemental Table 1: Listing of antibodies and antisera used in immunoblotting and immunocytochemical experiments. Supplemental Tables 2-7: Identification of cochlear proteins from sucrose gradients. The isolated proteins are tabulated in alphabetical order. For each protein, the accession number and the protein score defining the total fragment ions matching score (MS/MS matches only) is shown. Protein scores are derived from ion scores as a non-probabilistic basis for ranking protein hits. The ion score is -10·log(P), in which P is the probability that the observed match is a random event. We used stringent criteria for protein identification: a protein was designated as a “hit” only if it matched at least two distinct peptides with an ion score of at least 40. A single-peptide match was considered if the peptide had a good tandem mass spectrum and matched only a single protein. Because we used the non-redundant NCBI (NCBInr) Chordata database to search the tandem mass-spectrometric data, the accession numbers represent different species; when data from multiple species are used, that with the highest score is used. Also shown are the protein's mass, the peptide matches that define the total number of peptides matched to that protein, the protein coverage giving the percentage of peptide sequences matched to that protein, and the isoelectric point (PI). Supplemental Table 2: Identification of proteins from the sucrose pellet in the cochlea. Supplemental Table 3: Identification of proteins from the sucrose pellet in the retina. Supplemental Table 4: Identification of proteins from the harvested sucrose gradients between the 200-400 mM interface from cochlear material. Supplemental Table 5: Identification of proteins from the harvested sucrose gradients between the 400-600 mM interface from cochlear material. Supplemental Table 6: Identification of proteins from the harvested sucrose gradients between the 600-800 mM interface from cochlear material. Supplemental Table 7: Identification of proteins from the 47,000 x g pellet fraction from cochlear material. EM negative staining of the sucrose pellet fraction with the AD antibody indicating ribeye staining A B C D Scale bar: 100 nm EM negative staining of the vesicle fraction between the 200-400 mM sucrose layers using VAMP2 antibody A B C D E F Scale bar: 100 nm A. Coimmunoprecipitation with CtBP2 antibody: experiment I and experiment II (results in Table 2) eads P l b kDa kDa Ab flowthPreolulegthSupernaFtelonwt througChontro CTBP2 I Ab flowPtehlrleotuSguhpernaFtleonwtthroEulguhtion1Elution2Elution3Beads 120 120 100 ribeye 100 ribeye 80 80 60 60 50 50 CtBP2 CtBP2 50 CtBP1 40 myosin 9H 220 120 50 50 40 actin spectrin 30 40 20 30 60 50 40 B. Coimmunoprecipitation with AD antibody (Table 3 results) l o l t h r o n nt tr o n co kDa h as Pellet SupernateFlowthroWug Elution1 Elution2 Beads Elution cBeads 120 Ribeye 100 80 60 50 IgG CtBP2 120 100 80 PRPF39 60 IgG 50 A ribeye B syntaxin 1 C Merge B’ ribeye C’ GluR 2/3 D’ Merge Phalloidin Phalloidin D ribeye E SNAP25 F Merge Phalloidin E’ ribeye F’ vGlut3 G’ Merge Phalloidin G ribeye H VAMP2 I Merge Phalloidin H’ ribeye I’ Cav1.3 J’ Merge J ribeye K syntaxin 6 L Merge Phalloidin Phalloidin K’ ribeye L’ Otoferlin M’ Merge M ribeye N SNAP23 O Merge Phalloidin Phalloidin P ribeye Q VAMP7 R Merge Phalloidin N’ ribeye O’ rabphilin P’ Merge Phalloidin S ribeye T VAP-33 U Merge Phalloidin V ribeye W X Merge Phalloidin Q’ ribeye R’ munc18 S’ Merge Phalloidin Y ribeye Z NSF A’ Merge Phalloidin T’ ribeye U’ synapsin V’ Merge Phalloidin A ribeye B clathrin C Merge M ribeye N Sec13 O Merge Phalloidin Phalloidin P ribeye Q R Merge Phalloidin D ribeye E dynamin F Merge Phalloidin S ribeye T VCP U Merge Phalloidin G ribeye H SCAMP1 I Merge Phalloidin J endophilin K ribeye L Merge V ribeye W GAPDH X Merge Phalloidin Phalloidin A B ribeye C Merge Phalloidin D spectrin E CtBP2 F Merge Phalloidin G myosin H9 H CtBP2 I Merge Phalloidin J ribeye K L Merge Phalloidin M NF200 N CtBP2 O Merge Phalloidin P KIF3A Q CtBP2 R Merge Phalloidin Supplemental Table 1. List of antibodies and antisera. Antibodies Company and catalog number Concentrations Immunoblot Immunocytochemistry Otoferlin (R), HCS1 J.T. Corwin’s Laboratory NA 1:500 bassoon (M) Abcam ab76065 1:1000 1:500 NIPSNAP1 (R) Abcam ab67302 NA 1:200 SNAP25 (R) Abcam ab24737 1:1000 NA SNAP25 (m) Synaptic Systems 111 011 NA 1:200 VAMP1 (R) Abcam ab3346 1:2000 1:250 VAMP7 (M) Abcam ab36195 NA 1:100 clathrin heavy chain (M) Affinity Bioreagents MA1-065 1:500 1:250 VAMP2 (R) Assay Designs VAS-SV006F 1:2000 NA VAMP2 (M) Stressgen VAMSV014E NA 1:250 CASK (M) BD Transduction Laboratories 610782 1:500 NA CtBP1 (M) BD Transduction Laboratories 612042 1:2000 1:500 CtBP2 (M) BD Transduction Laboratories 612044 1:1000 1:500 dynamin (M) BD Transduction Laboratories 610246 1:1000 1:500 munc13 (M) BD Transduction Laboratories 610998 1:1000 NA munc18 (M) BD Transduction Laboratories 610337 1:1000 1:100 N-cadherin BD Transduction Laboratories 610920 1:2500 NA Rab3 (M) BD Transduction Laboratories 610380 1:2500 1:250 Rabphilin (M) BD Transduction Laboratories 610509 NA 1:250 RIM1 (M) BD Transduction Laboratories 610906 1:500 NA SCAMP1 (M) BD Transduction Laboratories 612087 NA 1:100 synapsin 1 (R) BD Transduction Laboratories 611392 1:10,000 NA Synapsin 1 (R) Hudspeth Laboratory NA 1:200 synaptotagmin1 (M) BD Transduction Laboratories 610434 1:500 NA syntaxin 4 (M) BD Transduction Laboratories 610439 1:10,000 NA syntaxin 6 (M) BD Transduction Laboratories 610635 1:2500 1:200 tomosyn (M) BD Transduction Laboratories 611297 1:500 NA VAP-33 (M) BD Transduction Laboratories 612180 1:2000 1:100 VCP97 (M) BD Transduction Laboratories 612182 NA 1:250 β-Catenin (M) BD Transduction Laboratories 610154 1:500 1:500 CaMKII (R) Cell Signalling 3361S NA 1:50 Complexin 1/2 (R) Chemicon International AB9288 1:1000 NA gluR 2 and 3 (R) Chemicon International AB 1506 NA 1:100 SV2 (M) Developmental Hybridoma Bank 1:1000 NA GAPDH (M) Millipore MAB374 1:1000 1:1000 GFAP (M) Novus Biologicals NB 300-141 1:1000 NA Sec13 (M) Novus Biologicals H00006396-A01 NA 1:100 β-COP (R) Pierce Biotechnology PA1-061 NA 1:100 CaBP4 (G) Santa Cruz Biotechnology sc-28096 1:500 1:500 synaptophysin 1 (R) Santa Cruz Biotechnology sc-9116 1:500 1:500 α-SNAP, β-SNAP (M) Santa Cruz Biotechnology sc-48349 1:250 1:100 actin (R) Sigma A 2066 1:500 1:250 KIF3A (R) Sigma K 3513 1:1000 1:200 Myosin H9 (R) Sigma M 8064 NA 1:500 NF160 (M) Sigma N 5264 1:1000 NA NF200 (R) Sigma N 4142 1:1000 1:250 NF68 (M) Sigma N 5139 1:1000 NA NSF (M) Sigma N 6038 1:2000 1:500 SNAP23 (R) Sigma Prestige Andtibodies HPA001214 1:2000 1:200 syntaxin 1 (M) Sigma S 0664 1:2000 1: 250 Synaptophysin (M) Sigma S 5768 Syntaxin 7 (R) Sigma S 4819 1:1000 NA α-tubulin (M) Sigma T 6074 1:2000 1:500 γ-tubulin (M) Sigma T 3559 1:10,000 NA Spectrin (M) Sigma Immunochemicals S1390 NA 1:200 PRPF39 (R) Sigma Prestige Antibodies HPA001176 NA 1:100 CSP (R) Stressgen VAP-SV003 1:5000 NA ERC2 (R) Synaptic Systems 143 103 1:1000 NA Piccolo (R) Synaptic Systems 142 002 1:2000 1:200 RIM2 (R) Synaptic Systems 140 103 1:1000 NA synaptojanin (R) Synaptic Systems 145 003 1:1000 NA VAMP3 (R) Synaptic Systems 104 103 1:2000 1:200 Cav1.3 (R) Teresa
Load more

Recommended publications
  • Supplementary Data
    Supplementary Data for Quantitative Changes in the Mitochondrial Proteome from Subjects with Mild Cognitive Impairment, Early Stage and Late Stage Alzheimer’s disease Table 1 - 112 unique, non-redundant proteins identified and quantified in at least two of the three analytical replicates for all three disease stages. Table 2 - MCI mitochondrial samples, Protein Summary Table 3 - MCI mitochondrial samples, Experiment 1 Table 4 - MCI mitochondrial samples, Experiment 2 Table 5 - MCI mitochondrial samples, Experiment 3 Table 6 - EAD Mitochondrial Study, Protein Summary Table 7 - EAD Mitochondrial Study, Experiment 1 Table 8 - EAD Mitochondrial Study, Experiment 2 Table 9 - EAD Mitochondrial Study, Experiment 3 Table 10 - LAD Mitochondrial Study, Protein Summary Table 11 - LAD Mitochondrial Study, Experiment 1 Table 12 - LAD Mitochondrial Study, Experiment 2 Table 13 - LAD Mitochondrial Study, Experiment 3 Supplemental Table 1. 112 unique, non-redundant proteins identified and quantified in at least two of the three analytical replicates for all three disease stages. Description Data MCI EAD LAD AATM_HUMAN (P00505) Aspartate aminotransferase, mitochondrial precursor (EC Mean 1.43 1.70 1.31 2.6.1.1) (Transaminase A) (Glutamate oxaloacetate transaminase 2) [MASS=47475] SEM 0.07 0.09 0.09 Count 3.00 3.00 3.00 ACON_HUMAN (Q99798) Aconitate hydratase, mitochondrial precursor (EC 4.2.1.3) Mean 1.24 1.61 1.19 (Citrate hydro-lyase) (Aconitase) [MASS=85425] SEM 0.05 0.17 0.18 Count 3.00 2.00 3.00 ACPM_HUMAN (O14561) Acyl carrier protein, mitochondrial
    [Show full text]
  • Proteomic Analyses Reveal a Role of Cytoplasmic Droplets As an Energy Source During Sperm Epididymal Maturation
    Proteomic analyses reveal a role of cytoplasmic droplets as an energy source during sperm epididymal maturation Shuiqiao Yuana,b, Huili Zhenga, Zhihong Zhengb, Wei Yana,1 aDepartment of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, NV, 89557; and bDepartment of Laboratory Animal Medicine, China Medical University, Shenyang, 110001, China Corresponding author. Email: [email protected] Supplemental Information contains one Figure (Figure S1), three Tables (Tables S1-S3) and two Videos (Videos S1 and S2) files. Figure S1. Scanning electron microscopic images of purified murine cytoplasmic droplets. Arrows point to indentations resembling the resealed defects at the detaching points when CDs come off the sperm flagella. Scale bar = 1µm Table S1 Mass spectrometry-based identifiaction of proteins highly enriched in murine cytoplasmic droplets. # MS/MS View:Identified Proteins (105) Accession Number Molecular Weight Protein Grouping Ambiguity Dot_1_1 Dot_2_1 Dot_3_1 Dot_4_1Dot_5_1 Dot_1_2 Dot_2_2 Dot_3_2 Dot_4_2 Dot_5_2 1 IPI:IPI00467457.3 Tax_Id=10090 Gene_Symbol=Ldhc L-lactate dehydrogenase C chain IPI00467457 36 kDa TRUE 91% 100% 100% 100% 100% 100% 100% 100% 100% 2 IPI:IPI00473320.2 Tax_Id=10090 Gene_Symbol=Actb Putative uncharacterized protein IPI00473320 42 kDa TRUE 75% 100% 100% 100% 100% 89% 76% 100% 100% 100% 3 IPI:IPI00224181.7 Tax_Id=10090 Gene_Symbol=Akr1b7 Aldose reductase-related protein 1 IPI00224181 36 kDa TRUE 100% 100% 76% 100% 100% 4 IPI:IPI00228633.7 Tax_Id=10090 Gene_Symbol=Gpi1 Glucose-6-phosphate
    [Show full text]
  • Establishing the Pathogenicity of Novel Mitochondrial DNA Sequence Variations: a Cell and Molecular Biology Approach
    Mafalda Rita Avó Bacalhau Establishing the Pathogenicity of Novel Mitochondrial DNA Sequence Variations: a Cell and Molecular Biology Approach Tese de doutoramento do Programa de Doutoramento em Ciências da Saúde, ramo de Ciências Biomédicas, orientada pela Professora Doutora Maria Manuela Monteiro Grazina e co-orientada pelo Professor Doutor Henrique Manuel Paixão dos Santos Girão e pela Professora Doutora Lee-Jun C. Wong e apresentada à Faculdade de Medicina da Universidade de Coimbra Julho 2017 Faculty of Medicine Establishing the pathogenicity of novel mitochondrial DNA sequence variations: a cell and molecular biology approach Mafalda Rita Avó Bacalhau Tese de doutoramento do programa em Ciências da Saúde, ramo de Ciências Biomédicas, realizada sob a orientação científica da Professora Doutora Maria Manuela Monteiro Grazina; e co-orientação do Professor Doutor Henrique Manuel Paixão dos Santos Girão e da Professora Doutora Lee-Jun C. Wong, apresentada à Faculdade de Medicina da Universidade de Coimbra. Julho, 2017 Copyright© Mafalda Bacalhau e Manuela Grazina, 2017 Esta cópia da tese é fornecida na condição de que quem a consulta reconhece que os direitos de autor são pertença do autor da tese e do orientador científico e que nenhuma citação ou informação obtida a partir dela pode ser publicada sem a referência apropriada e autorização. This copy of the thesis has been supplied on the condition that anyone who consults it recognizes that its copyright belongs to its author and scientific supervisor and that no quotation from the
    [Show full text]
  • MYH9-Related Platelet Disorders
    Reprinted with permission from Thieme Medical Publishers (Semin Thromb Hemost 2009;35:189-203) Homepage at www.thieme.com MYH9-Related Platelet Disorders Karina Althaus, M.D.,1 and Andreas Greinacher, M.D.1 ABSTRACT Myosin heavy chain 9 (MYH9)-related platelet disorders belong to the group of inherited thrombocytopenias. The MYH9 gene encodes the nonmuscle myosin heavy chain IIA (NMMHC-IIA), a cytoskeletal contractile protein. Several mutations in the MYH9 gene lead to premature release of platelets from the bone marrow, macro- thrombocytopenia, and cytoplasmic inclusion bodies within leukocytes. Four overlapping syndromes, known as May-Hegglin anomaly, Epstein syndrome, Fechtner syndrome, and Sebastian platelet syndrome, describe different clinical manifestations of MYH9 gene mutations. Macrothrombocytopenia is present in all affected individuals, whereas only some develop additional clinical manifestations such as renal failure, hearing loss, and presenile cataracts. The bleeding tendency is usually moderate, with menorrhagia and easy bruising being most frequent. The biggest risk for the individual is inappropriate treatment due to misdiagnosis of chronic autoimmune thrombocytopenia. To date, 31 mutations of the MYH9 gene leading to macrothrombocytopenia have been identified, of which the upstream mutations up to amino acid 1400 are more likely associated with syndromic manifestations than the downstream mutations. This review provides a short history of MYH9-related disorders, summarizes the clinical and laboratory character- istics, describes a diagnostic algorithm, presents recent results of animal models, and discusses aspects of therapeutic management. KEYWORDS: MYH9 gene, nonmuscle myosin IIA, May-Hegglin anomaly, Epstein syndrome, Fechtner syndrome, Sebastian platelet syndrome, macrothrombocytopenia The correct diagnosis of hereditary chronic as isolated platelet count reductions or as part of thrombocytopenias is important for planning appropri- more complex clinical syndromes.
    [Show full text]
  • Table 1 Gene Name Increased Or Decreased in LTD
    Table_1 gene_name increased or decreased in LTD protein_id description keep_supernatant keep_pellet comparison sample hit_annotation_methodpvalue fdr hit hit_annotation 2010300C02RIK increased E9Q3M9 Protein 2010300C02Rik OS=Mus musculus GN=2010300C02Rik PE=1 SV=1 TRUE TRUE NMDA - control pellet fdrtool 0,074667 0,584087 FALSE trend 2310035C23RIK|KIAA1468increased A0A087WSS1|E9QM90|Q148V7|Q148V7-2 Protein 2310035C23Rik OS=Mus musculus GN=2310035C23Rik PE=1 SV=1|Protein 2310035C23Rik OS=Mus musculusTRUE GN=2310035C23Rik PE=1 SV=2|LisH FALSE domain NMDAand HEAT - control repeat-containing protein KIAA1468 supernatant OS=Mus musculus GN=Kiaa1468 fdrtoolPE=1 SV=1|Isoform 0,080056 2 of 0,589077LisH domain FALSEand HEAT trend repeat-containing protein KIAA1468 OS=Mus musculus GN=Kiaa1468 ABR increased E9PUE7|Q5SSL4|Q5SSL4-2|Q5SSL4-3|Q5SSL4-4 Active breakpoint cluster region-related protein OS=Mus musculus GN=Abr PE=1 SV=1|Isoform 2 of Active breakpointTRUE cluster region-related protein TRUE OS=Mus musculus NMDA GN=Abr|Isoform - control 3 of Active breakpoint supernatant cluster region-related protein OS=Mus fdrtool musculus 0,08128 GN=Abr|Isoform 0,592743 4 of Active FALSE breakpoint trend cluster region-related protein OS=Mus musculus GN=Abr ADAM22 increased D3YUP9|Q9R1V6|Q9R1V6-10|Q9R1V6-11|Q9R1V6-12|Q9R1V6-13|Q9R1V6-14|Q9R1V6-15|Q9R1V6-17|Q9R1V6-4|Q9R1V6-5|Q9R1V6-6|Q9R1V6-7|Q9R1V6-8Disintegrin and metalloproteinase domain-containing protein 22 OS=Mus musculus GN=Adam22 PE=1 SV=1|DisintegrinFALSE and metalloproteinase domain-containing TRUE
    [Show full text]
  • Molecular Profile of Tumor-Specific CD8+ T Cell Hypofunction in a Transplantable Murine Cancer Model
    Downloaded from http://www.jimmunol.org/ by guest on September 25, 2021 T + is online at: average * The Journal of Immunology , 34 of which you can access for free at: 2016; 197:1477-1488; Prepublished online 1 July from submission to initial decision 4 weeks from acceptance to publication 2016; doi: 10.4049/jimmunol.1600589 http://www.jimmunol.org/content/197/4/1477 Molecular Profile of Tumor-Specific CD8 Cell Hypofunction in a Transplantable Murine Cancer Model Katherine A. Waugh, Sonia M. Leach, Brandon L. Moore, Tullia C. Bruno, Jonathan D. Buhrman and Jill E. Slansky J Immunol cites 95 articles Submit online. Every submission reviewed by practicing scientists ? is published twice each month by Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts http://jimmunol.org/subscription Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html http://www.jimmunol.org/content/suppl/2016/07/01/jimmunol.160058 9.DCSupplemental This article http://www.jimmunol.org/content/197/4/1477.full#ref-list-1 Information about subscribing to The JI No Triage! Fast Publication! Rapid Reviews! 30 days* Why • • • Material References Permissions Email Alerts Subscription Supplementary The Journal of Immunology 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. This information is current as of September 25, 2021. The Journal of Immunology Molecular Profile of Tumor-Specific CD8+ T Cell Hypofunction in a Transplantable Murine Cancer Model Katherine A.
    [Show full text]
  • An Animal Model with a Cardiomyocyte-Specific Deletion of Estrogen Receptor Alpha: Functional, Metabolic, and Differential Netwo
    Washington University School of Medicine Digital Commons@Becker Open Access Publications 2014 An animal model with a cardiomyocyte-specific deletion of estrogen receptor alpha: Functional, metabolic, and differential network analysis Sriram Devanathan Washington University School of Medicine in St. Louis Timothy Whitehead Washington University School of Medicine in St. Louis George G. Schweitzer Washington University School of Medicine in St. Louis Nicole Fettig Washington University School of Medicine in St. Louis Attila Kovacs Washington University School of Medicine in St. Louis See next page for additional authors Follow this and additional works at: https://digitalcommons.wustl.edu/open_access_pubs Recommended Citation Devanathan, Sriram; Whitehead, Timothy; Schweitzer, George G.; Fettig, Nicole; Kovacs, Attila; Korach, Kenneth S.; Finck, Brian N.; and Shoghi, Kooresh I., ,"An animal model with a cardiomyocyte-specific deletion of estrogen receptor alpha: Functional, metabolic, and differential network analysis." PLoS One.9,7. e101900. (2014). https://digitalcommons.wustl.edu/open_access_pubs/3326 This Open Access Publication is brought to you for free and open access by Digital Commons@Becker. It has been accepted for inclusion in Open Access Publications by an authorized administrator of Digital Commons@Becker. For more information, please contact [email protected]. Authors Sriram Devanathan, Timothy Whitehead, George G. Schweitzer, Nicole Fettig, Attila Kovacs, Kenneth S. Korach, Brian N. Finck, and Kooresh I. Shoghi This open access publication is available at Digital Commons@Becker: https://digitalcommons.wustl.edu/open_access_pubs/3326 An Animal Model with a Cardiomyocyte-Specific Deletion of Estrogen Receptor Alpha: Functional, Metabolic, and Differential Network Analysis Sriram Devanathan1, Timothy Whitehead1, George G. Schweitzer2, Nicole Fettig1, Attila Kovacs3, Kenneth S.
    [Show full text]
  • Functions of Vertebrate Ferlins
    cells Review Functions of Vertebrate Ferlins Anna V. Bulankina 1 and Sven Thoms 2,* 1 Department of Internal Medicine 1, Goethe University Hospital Frankfurt, 60590 Frankfurt, Germany; [email protected] 2 Department of Child and Adolescent Health, University Medical Center Göttingen, 37075 Göttingen, Germany * Correspondence: [email protected] Received: 27 January 2020; Accepted: 20 February 2020; Published: 25 February 2020 Abstract: Ferlins are multiple-C2-domain proteins involved in Ca2+-triggered membrane dynamics within the secretory, endocytic and lysosomal pathways. In bony vertebrates there are six ferlin genes encoding, in humans, dysferlin, otoferlin, myoferlin, Fer1L5 and 6 and the long noncoding RNA Fer1L4. Mutations in DYSF (dysferlin) can cause a range of muscle diseases with various clinical manifestations collectively known as dysferlinopathies, including limb-girdle muscular dystrophy type 2B (LGMD2B) and Miyoshi myopathy. A mutation in MYOF (myoferlin) was linked to a muscular dystrophy accompanied by cardiomyopathy. Mutations in OTOF (otoferlin) can be the cause of nonsyndromic deafness DFNB9. Dysregulated expression of any human ferlin may be associated with development of cancer. This review provides a detailed description of functions of the vertebrate ferlins with a focus on muscle ferlins and discusses the mechanisms leading to disease development. Keywords: dysferlin; myoferlin; otoferlin; C2 domain; calcium-sensor; muscular dystrophy; dysferlinopathy; limb girdle muscular dystrophy type 2B (LGMD2B), membrane repair; T-tubule system; DFNB9 1. Introduction Ferlins belong to the superfamily of proteins with multiple C2 domains (MC2D) that share common functions in tethering membrane-bound organelles or recruiting proteins to cellular membranes. Ferlins are described as calcium ions (Ca2+)-sensors for vesicular trafficking capable of sculpturing membranes [1–3].
    [Show full text]
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • HESN CROI Poster
    HIV-Exposed Seronegative MSM express antiproteases with novel antiviral activity Laura Romas1,2, Klara Hasselrot3, Carolina Hererra4, Garrett Westmacott5, Francis Plummer5,1, T. Blake Ball2,1, Kristina Broliden3, Adam Burgener2,1,3 1. Dept. of Med. Microbiology, University of Manitoba, CAN 2. National HIV and Retrovirology Laboratory, JC Wilt Infectioius Disease Research Centre, Public Health Agency of Canada; Poster #287 3. Dept. of Medicine Solna, Center for Molecular Medicine, Karolinska Institutet, SWE; 4. Imperial College of London, UK; 5. National Microbiology Laboratory, Public Health Agency of Canada, CAN INTRODUCTION RESULTS SUMMARY •The risk of HIV acquisition through the rectum is signicantly higher than other sites of mucosal exposure, which contributes to disproportionate rates of infection A in at-risk men who have sex with men (MSM) practicing unprotected receptive 1 anal intercourse (URAI). •HIV susceptibility at the rectal mucosa a result of a thin columnar epithlia, pres- ence of activated T cells within the submucosa, and a tighly associated lymphatic Antprotease 2 Antprotease 1 system for easy viral disseminatio(Reviewed in 1). •However, the immunobiology of rectal mucosa, and factors which aect HIV sus- ceptibility are not well understood and represents a major barrier to the develop- ment of prevention technologies. Our recent proteomic analysis of rectal mucosa secretions suggests that this uid contains hundreds of innate factors important for host defense, and is immunologically distinct from other mucosal compart- ments and sites of HIV exposure1. •Study of HIV-Exposed Seronegative (HESN) individuals have shown altered mu- cosal immune responses in cervical, salivary and foreskin secretions associated with reduced HIV-susceptibility2-4.
    [Show full text]
  • Cellular and Molecular Signatures in the Disease Tissue of Early
    Cellular and Molecular Signatures in the Disease Tissue of Early Rheumatoid Arthritis Stratify Clinical Response to csDMARD-Therapy and Predict Radiographic Progression Frances Humby1,* Myles Lewis1,* Nandhini Ramamoorthi2, Jason Hackney3, Michael Barnes1, Michele Bombardieri1, Francesca Setiadi2, Stephen Kelly1, Fabiola Bene1, Maria di Cicco1, Sudeh Riahi1, Vidalba Rocher-Ros1, Nora Ng1, Ilias Lazorou1, Rebecca E. Hands1, Desiree van der Heijde4, Robert Landewé5, Annette van der Helm-van Mil4, Alberto Cauli6, Iain B. McInnes7, Christopher D. Buckley8, Ernest Choy9, Peter Taylor10, Michael J. Townsend2 & Costantino Pitzalis1 1Centre for Experimental Medicine and Rheumatology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, Charterhouse Square, London EC1M 6BQ, UK. Departments of 2Biomarker Discovery OMNI, 3Bioinformatics and Computational Biology, Genentech Research and Early Development, South San Francisco, California 94080 USA 4Department of Rheumatology, Leiden University Medical Center, The Netherlands 5Department of Clinical Immunology & Rheumatology, Amsterdam Rheumatology & Immunology Center, Amsterdam, The Netherlands 6Rheumatology Unit, Department of Medical Sciences, Policlinico of the University of Cagliari, Cagliari, Italy 7Institute of Infection, Immunity and Inflammation, University of Glasgow, Glasgow G12 8TA, UK 8Rheumatology Research Group, Institute of Inflammation and Ageing (IIA), University of Birmingham, Birmingham B15 2WB, UK 9Institute of
    [Show full text]
  • Serum Albumin OS=Homo Sapiens
    Protein Name Cluster of Glial fibrillary acidic protein OS=Homo sapiens GN=GFAP PE=1 SV=1 (P14136) Serum albumin OS=Homo sapiens GN=ALB PE=1 SV=2 Cluster of Isoform 3 of Plectin OS=Homo sapiens GN=PLEC (Q15149-3) Cluster of Hemoglobin subunit beta OS=Homo sapiens GN=HBB PE=1 SV=2 (P68871) Vimentin OS=Homo sapiens GN=VIM PE=1 SV=4 Cluster of Tubulin beta-3 chain OS=Homo sapiens GN=TUBB3 PE=1 SV=2 (Q13509) Cluster of Actin, cytoplasmic 1 OS=Homo sapiens GN=ACTB PE=1 SV=1 (P60709) Cluster of Tubulin alpha-1B chain OS=Homo sapiens GN=TUBA1B PE=1 SV=1 (P68363) Cluster of Isoform 2 of Spectrin alpha chain, non-erythrocytic 1 OS=Homo sapiens GN=SPTAN1 (Q13813-2) Hemoglobin subunit alpha OS=Homo sapiens GN=HBA1 PE=1 SV=2 Cluster of Spectrin beta chain, non-erythrocytic 1 OS=Homo sapiens GN=SPTBN1 PE=1 SV=2 (Q01082) Cluster of Pyruvate kinase isozymes M1/M2 OS=Homo sapiens GN=PKM PE=1 SV=4 (P14618) Glyceraldehyde-3-phosphate dehydrogenase OS=Homo sapiens GN=GAPDH PE=1 SV=3 Clathrin heavy chain 1 OS=Homo sapiens GN=CLTC PE=1 SV=5 Filamin-A OS=Homo sapiens GN=FLNA PE=1 SV=4 Cytoplasmic dynein 1 heavy chain 1 OS=Homo sapiens GN=DYNC1H1 PE=1 SV=5 Cluster of ATPase, Na+/K+ transporting, alpha 2 (+) polypeptide OS=Homo sapiens GN=ATP1A2 PE=3 SV=1 (B1AKY9) Fibrinogen beta chain OS=Homo sapiens GN=FGB PE=1 SV=2 Fibrinogen alpha chain OS=Homo sapiens GN=FGA PE=1 SV=2 Dihydropyrimidinase-related protein 2 OS=Homo sapiens GN=DPYSL2 PE=1 SV=1 Cluster of Alpha-actinin-1 OS=Homo sapiens GN=ACTN1 PE=1 SV=2 (P12814) 60 kDa heat shock protein, mitochondrial OS=Homo
    [Show full text]