DOCSLIB.ORG

SUPT6H Controls Estrogen Receptor Activity and Cellular Differentiation by Multiple Epigenomic Mechanisms

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
SUPT6H Controls Estrogen Receptor Activity and Cellular Differentiation by Multiple Epigenomic Mechanisms Oncogene (2015) 34, 465–473 & 2015 Macmillan Publishers Limited All rights reserved 0950-9232/15 www.nature.com/onc ORIGINAL ARTICLE SUPT6H controls estrogen receptor activity and cellular differentiation by multiple epigenomic mechanisms U Bedi1,2, AH Scheel3,4, M Hennion2, Y Begus-Nahrmann2,JRu¨ schoff3 and SA Johnsen1,2 The estrogen receptor alpha (ERa) is the central transcriptional regulator of ductal mammary epithelial lineage specification and is an important prognostic marker in human breast cancer. Although antiestrogen therapies are initially highly effective at treating ERa-positive tumors, a large number of tumors progress to a refractory, more poorly differentiated phenotype accompanied by reduced survival. A better understanding of the molecular mechanisms involved in the progression from estrogen-dependent to hormone-resistant breast cancer may uncover new targets for treatment and the discovery of new predictive markers. Recent studies have uncovered an important role for transcriptional elongation and chromatin modifications in controlling ERa activity and estrogen responsiveness. The human Suppressor of Ty Homologue-6 (SUPT6H) is a histone chaperone that links transcriptional elongation to changes in chromatin structure. We show that SUPT6H is required for estrogen-regulated transcription and the maintenance of chromatin structure in breast cancer cells, possibly in part through interaction with RNF40 and regulation of histone H2B monoubiquitination (H2Bub1). Moreover, we demonstrate that SUPT6H protein levels decrease with malignancy in breast cancer. Consistently, SUPT6H, similar to H2Bub1, is required for cellular differentiation and suppression of the repressive histone mark H3K27me3 on lineage-specific genes. Together, these data identify SUPT6H as a new epigenetic regulator of ERa activity and cellular differentiation. Oncogene (2015) 34, 465–473; doi:10.1038/onc.2013.558; published online 20 January 2014 Keywords: estrogen receptor; breast cancer; differentiation; chromatin INTRODUCTION enzymes, changes in histone modifications, chromatin remodeling The estrogen receptor a (ERa) is one of the key transcriptional and significant changes in overall chromatin organization.10–13 regulators of proliferation and differentiation in the mammary These changes appear to be required for the efficient expression epithelium.1 Approximately two-thirds of human breast cancers of estrogen-regulated genes and may represent new potential express ERa.ERa expression is associated with a more therapeutic targets.14 In previous studies, we uncovered a tumor differentiated luminal tumor phenotype and overall better suppressor function for H2Bub115–17 where its levels decrease patient survival compared with ERa-negative tumors.2,3 Thus, during the malignant progression of breast cancer. Interestingly, ERa expression is an important prognostic marker and is although H2Bub1 is required for ERa-regulated gene transcription, predictive for tumor response to antiestrogen treatment. Despite instead of leading to impaired cell proliferation, a loss of H2Bub1 an initial positive response, roughly one-third of ERa-positive instead led to the estrogen-independent growth of ERa-positive tumors becomes refractory to antihormone therapy and develops MCF7 breast cancer cells,15 possibly implicating a loss of H2Bub1 estrogen-independence. Therefore, an increased understanding in the transition from an estrogen-dependent to hormone- of the molecular mechanisms controlling ERa-mediated tran- independent growth of breast cancer. scriptional regulation may help to uncover new molecular targets Consistent with the inverse correlation between H2Bub1 and which may be utilized to more effectively treat and eradicate differentiation status in breast cancer, H2Bub1 levels increase ERa-positive tumors.4 during the differentiation of various types of somatic and Recent studies have demonstrated that one of the essential pluripotent stem cells.18–20 Furthermore, a loss of H2Bub1 results transcriptional regulatory steps controlled by ERa is transcriptional in impaired differentiation in diverse stem cell systems and elongation.5 ERa interacts with the Positive Transcription organisms indicating that H2Bub1 levels are intimately connected Elongation Factor-b complex6 that promotes transcriptional to differentiation status in both normal and tumor cells. elongation in part by phosphorylating Ser2 (pSer2) within the Mechanistically, H2Bub1 appears to be associated with tran- carboxy-terminal domain of RNA Polymerase II (RNAPII).7 HEXIM1, scriptional elongation where it is localized within the transcribed a negative regulator of Positive Transcription Elongation Factor-b region of active genes and its presence is associated with gene activity also interacts with ERa8 and its overexpression leads to a expression levels.21 Consistently, H2B monoubiquitination is tamoxifen-resistant phenotype in breast cancer cells.9 dependent upon the activity of Positive Transcription Elongation ERa-regulated transcription is a highly dynamic process which is Factor-b and phosphorylation of Ser2,22,23 where the obligate associated with the recruitment of multiple histone-modifying heterodimeric H2B ubiquitin ligase complex RNF20/40 is linked to 1Center of Experimental Medicine, Department of Tumor Biology, University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany; 2Institute of Molecular Oncology, University Medical Center Go¨ttingen, Go¨ttingen, Germany; 3Institute of Pathology Nordhessen, Kassel, Germany and 4Center for Pathology and Forensic Medicine, Department of Pathology, University Medical Center Go¨ttingen, Go¨ttingen, Germany. Correspondence: Professor SA Johnsen, Center of Experimental Medicine, Department of Tumor Biology, University Medical Center Hamburg-Eppendorf (UKE), Hamburg 20251, Germany. E-mail: [email protected] Received 15 July 2013; revised 21 November 2013; accepted 22 November 2013; published online 20 January 2014 SUPT6H controls estrogen receptor activity and cellular differentiation U Bedi et al 466 elongating RNAPII via the WW domain containing adapter with RESULTS coiled-coil (WAC) protein that binds directly to pSer2.24 Similar to SUPT6H is required for ERa activity WAC, the histone chaperone Suppressor of Ty Homologue-6 Given the importance of transcriptional elongation in the (SUPT6H) binds to the elongating Ser2 phosphorylated form of 25,26 regulation of ERa activity, we hypothesized that the histone RNAPII. As the yeast Suppressor of Ty-6 (Spt6) protein was chaperone and transcriptional elongation regulatory protein, shown to be required for transcriptional regulation by human ERa 27 SUPT6H may also be required for estrogen-regulated transcription. in yeast, we hypothesized that SUPT6H may also regulate ERa In order to test this hypothesis, we compared the effects of small activity and influence H2Bub1 during cellular differentiation and interfering RNA-mediated knockdown of SUPT6H to those of the tumor progression. In order to address this hypothesis, we pure ERa antagonist, ICI 1 82 780 on the induction of several investigated the function of SUPT6H in ERa-positive MCF7 and estrogen-regulated genes in the ERa-positive luminal breast T47D breast cancer cells and demonstrate that SUPT6H is required cancer cells, MCF7 and T47D. Indeed, the induction of the for estrogen-regulated gene transcription and the maintenance of investigated direct ERa target genes (CXCL12, GREB1 and PGR) was chromatin structure. We show that these effects are linked to similarly decreased following SUPT6H knockdown or ICI 1 82 780 H2Bub1, where SUPT6H and H2Bub1 levels correlate with the treatment in both the cell lines (Figures 1a and b) while the differentiation status of primary breast tumors and similarly expression of TTP and DSP and ERa expression were not affected regulate cellular differentiation via removal of the repressive by either SUPT6H knockdown or ICI 1 82 780 treatment mark H3K27me3. These findings identify SUPT6H as a central (Supplementary Figures S1a–d). Furthermore, since SUPT6H regulator of ERa activity and cellular differentiation status and displays histone chaperone activity and may therefore promote suggest that alterations affecting transcriptional elongation and the opening of chromatin and binding of ERa, we next tested associated changes in chromatin structure may have a central role whether its depletion affects ERa recruitment to chromatin at in malignant progression. Estrogen Response Elements (ERE) on two well characterized Figure 1. SUPT6H knockdown decreases estrogen-induced gene expression by reducing ERa recruitment to target genes. (a, b) MCF7 and T47D cells were transfected with either control or SUPT6H small interfering RNA (siRNAs) or treated with ICI 1 82 780, grown for 24 h before switching to hormone-deprived medium and grown for another 24 h. Cells were then stimulated with 10 nmol/l 17b-estradiol (E2) for 6 h and the expression levels of CXCL12, GREB1, PGR and SUPT6H were analyzed by quantitative PCR (qPCR). Gene expression levels were normalized to 18S ribosomal RNA, graphed relative to the control sample and expressed as ‘Relative mRNA Expression’; mean values þ s.d., n ¼ 3. (c, d) Decreased ERa and SUPT6H recruitment to distal EREs of GREB1 and PGR upon SUPT6H knockdown. MCF7 cells were transfected and cultured as in (a), except that cells were stimulated with 10 nmol/l 17b-estradiol (E2) for 2 h. ChIP samples were normalized to input samples and expressed as ‘% Input’; mean values þ s.d., n ¼ 3. The
Load more

Recommended publications
  • FACT Sets a Barrier for Cell Fate Reprogramming in C. Elegans and Human
    bioRxiv preprint doi: https://doi.org/10.1101/185116; this version posted September 6, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. FACT sets a barrier for cell fate reprogramming in C. elegans and Human Authors: Ena Kolundzic1, Andreas Ofenbauer1, Bora Uyar1, Anne Sommermeier1,2, Stefanie Seelk1, Mei He1, Gülkiz Baytek1, Altuna Akalin1, Sebastian Diecke1,3*, Scott A. Lacadie1,3*, Baris Tursun1* Affiliations: 1Berlin Institute for Medical Systems Biology, Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany; 2Department of Biology, Humboldt University, Berlin, Germany; 3 Berlin Institute of Health, Berlin, Germany *Correspondence to: [email protected] (BT), [email protected] (SL), [email protected] (SD) Lead Contact: [email protected] (BT) 1 bioRxiv preprint doi: https://doi.org/10.1101/185116; this version posted September 6, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Summary: The chromatin regulator FACT (Facilitates Chromatin Transcription) is essential for ensuring stable gene expression by promoting transcription. In a genetic screen using C. elegans we identified that FACT maintains cell identities and acts as a barrier for transcription factor-mediated cell fate reprogramming. Strikingly, FACT’s role as a reprogramming barrier is conserved in humans as we show that FACT depletion enhances reprogramming of fibroblasts into stem cells and neurons. Such activity of FACT is unexpected since known reprogramming barriers typically repress gene expression by silencing chromatin.
    [Show full text]
  • Genomic Correlates of Relationship QTL Involved in Fore- Versus Hind Limb Divergence in Mice
    Loyola University Chicago Loyola eCommons Biology: Faculty Publications and Other Works Faculty Publications 2013 Genomic Correlates of Relationship QTL Involved in Fore- Versus Hind Limb Divergence in Mice Mihaela Palicev Gunter P. Wagner James P. Noonan Benedikt Hallgrimsson James M. Cheverud Loyola University Chicago, [email protected] Follow this and additional works at: https://ecommons.luc.edu/biology_facpubs Part of the Biology Commons Recommended Citation Palicev, M, GP Wagner, JP Noonan, B Hallgrimsson, and JM Cheverud. "Genomic Correlates of Relationship QTL Involved in Fore- Versus Hind Limb Divergence in Mice." Genome Biology and Evolution 5(10), 2013. This Article is brought to you for free and open access by the Faculty Publications at Loyola eCommons. It has been accepted for inclusion in Biology: Faculty Publications and Other Works by an authorized administrator of Loyola eCommons. For more information, please contact [email protected]. This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License. © Palicev et al., 2013. GBE Genomic Correlates of Relationship QTL Involved in Fore- versus Hind Limb Divergence in Mice Mihaela Pavlicev1,2,*, Gu¨ nter P. Wagner3, James P. Noonan4, Benedikt Hallgrı´msson5,and James M. Cheverud6 1Konrad Lorenz Institute for Evolution and Cognition Research, Altenberg, Austria 2Department of Pediatrics, Cincinnati Children‘s Hospital Medical Center, Cincinnati, Ohio 3Yale Systems Biology Institute and Department of Ecology and Evolutionary Biology, Yale University 4Department of Genetics, Yale University School of Medicine 5Department of Cell Biology and Anatomy, The McCaig Institute for Bone and Joint Health and the Alberta Children’s Hospital Research Institute for Child and Maternal Health, University of Calgary, Calgary, Canada 6Department of Anatomy and Neurobiology, Washington University *Corresponding author: E-mail: [email protected].
    [Show full text]
  • Differential Expression of Multiple Disease-Related Protein Groups
    brain sciences Article Differential Expression of Multiple Disease-Related Protein Groups Induced by Valproic Acid in Human SH-SY5Y Neuroblastoma Cells 1,2, 1, 1 1 Tsung-Ming Hu y, Hsiang-Sheng Chung y, Lieh-Yung Ping , Shih-Hsin Hsu , Hsin-Yao Tsai 1, Shaw-Ji Chen 3,4 and Min-Chih Cheng 1,* 1 Department of Psychiatry, Yuli Branch, Taipei Veterans General Hospital, Hualien 98142, Taiwan; [email protected] (T.-M.H.); [email protected] (H.-S.C.); [email protected] (L.-Y.P.); fi[email protected] (S.-H.H.); [email protected] (H.-Y.T.) 2 Department of Future Studies and LOHAS Industry, Fo Guang University, Jiaosi, Yilan County 26247, Taiwan 3 Department of Psychiatry, Mackay Medical College, New Taipei City 25245, Taiwan; [email protected] 4 Department of Psychiatry, Taitung Mackay Memorial Hospital, Taitung County 95064, Taiwan * Correspondence: [email protected]; Tel.: +886-3888-3141 (ext. 475) These authors contributed equally to this work. y Received: 10 July 2020; Accepted: 8 August 2020; Published: 12 August 2020 Abstract: Valproic acid (VPA) is a multifunctional medication used for the treatment of epilepsy, mania associated with bipolar disorder, and migraine. The pharmacological effects of VPA involve a variety of neurotransmitter and cell signaling systems, but the molecular mechanisms underlying its clinical efficacy is to date largely unknown. In this study, we used the isobaric tags for relative and absolute quantitation shotgun proteomic analysis to screen differentially expressed proteins in VPA-treated SH-SY5Y cells. We identified changes in the expression levels of multiple proteins involved in Alzheimer’s disease, Parkinson’s disease, chromatin remodeling, controlling gene expression via the vitamin D receptor, ribosome biogenesis, ubiquitin-mediated proteolysis, and the mitochondrial oxidative phosphorylation and electron transport chain.
    [Show full text]
  • Prenatal Diagnosis of Maternal Partial Trisomy 9P23p24.3 and 14Q11.2Q21.3 in a Fetus: a Case Report J
    Wu et al. Molecular Cytogenetics (2020) 13:6 https://doi.org/10.1186/s13039-020-0473-x CASE REPORT Open Access Prenatal diagnosis of maternal partial trisomy 9p23p24.3 and 14q11.2q21.3 in a fetus: a case report J. B. Wu1†, J. Sha1†, J. F. Zhai1*, Y. Liu2 and B. Zhang1 Abstract Objective: This study aimed to report a fetus with maternal partial trisomy 9p and 14q and the phenotype detected in ultrasound. Methods: The chromosome rearrangements in the fetus were characterized by G-banding and chromosome microarray analysis based on single nucleotide polymorphism (SNP) array of cultured amniocytes and compared with the parents’ karyotypes. Results: The fetal abnormal karyotype was 47,XY,+der(14)(9;14)(p23;q22). The SNP array revealed a duplicate 11.8- Mb 9p23-p24.3 fragment and a duplicate 29.6-Mb 14q11.2-q21.3 fragment. The peripheral blood karyotype of the mother was 46,XX,t(9;14)(p23;q22), while the father’s was normal at the level of 300~400 bands. A high-resolution karyotype analysis conformed the same abnormality of the mother at the level of 550~650 bands. These results indicated that the fetal chromosomal abnormality probably derived from the mother. The fetal nuchal translucency thickness was 3.5 mm, and the fetal heart was detected with around 1.0-mm ventricular defect by the ultrasound examination at 12-week gestation. The couple decided to terminate the pregnancy. They opted for in vitro fertilization and embryo transfer for the fourth pregnancy, which was successful. Conclusions: The SNP array combined with cytogenetic analysis was particularly effective in identifying abnormal chromosomal rearrangements.
    [Show full text]
  • Histone Monoubiquitination in Chromatin Remodelling: Focus on the Histone H2B Interactome and Cancer
    cancers Review Histone Monoubiquitination in Chromatin Remodelling: Focus on the Histone H2B Interactome and Cancer Deborah J. Marsh 1,2,* , Yue Ma 1 and Kristie-Ann Dickson 1 1 Translational Oncology Group, Faculty of Science, School of Life Sciences, University of Technology Sydney, Ultimo, NSW 2007, Australia; [email protected] (Y.M.); [email protected] (K.-A.D.) 2 Kolling Institute, Faculty of Medicine and Health, Northern Clinical School, University of Sydney, Camperdown, NSW 2006, Australia * Correspondence: [email protected]; Tel.: +61-2-9514-7574 Received: 17 October 2020; Accepted: 17 November 2020; Published: 20 November 2020 Simple Summary: Post-translational modifications (PTM) of histone tails represent epigenomic regulation of the chromatin landscape, influencing gene expression and the response to DNA damage. This review focusses on cancer-associated roles of ubiquitin as a histone PTM, specifically in conjunction with an E3 ubiquitin ligase cascade that results in the addition of a single ubiquitin (monoubiquitination) to histone H2B at lysine 120 (H2Bub1). H2Bub1 has roles in chromatin accessibility important for transcriptional elongation, the DNA damage response, cellular proliferation and developmental transitions, including in stem cell plasticity. It has been implicated in inflammation and tumour progression, with examples of its loss associated with a worse prognosis for patients with some cancers. Many factors involved in the H2Bub1 interactome are well known cancer-associated proteins, including p53, BRCA1 and components of the SWI/SNF remodelling complex. Increased knowledge of H2Bub1 and its interactome offers new opportunities for therapeutic targeting of malignancy. Abstract: Chromatin remodelling is a major mechanism by which cells control fundamental processes including gene expression, the DNA damage response (DDR) and ensuring the genomic plasticity required by stem cells to enable differentiation.
    [Show full text]
  • Characterizing Genomic Duplication in Autism Spectrum Disorder by Edward James Higginbotham a Thesis Submitted in Conformity
    Characterizing Genomic Duplication in Autism Spectrum Disorder by Edward James Higginbotham A thesis submitted in conformity with the requirements for the degree of Master of Science Graduate Department of Molecular Genetics University of Toronto © Copyright by Edward James Higginbotham 2020 i Abstract Characterizing Genomic Duplication in Autism Spectrum Disorder Edward James Higginbotham Master of Science Graduate Department of Molecular Genetics University of Toronto 2020 Duplication, the gain of additional copies of genomic material relative to its ancestral diploid state is yet to achieve full appreciation for its role in human traits and disease. Challenges include accurately genotyping, annotating, and characterizing the properties of duplications, and resolving duplication mechanisms. Whole genome sequencing, in principle, should enable accurate detection of duplications in a single experiment. This thesis makes use of the technology to catalogue disease relevant duplications in the genomes of 2,739 individuals with Autism Spectrum Disorder (ASD) who enrolled in the Autism Speaks MSSNG Project. Fine-mapping the breakpoint junctions of 259 ASD-relevant duplications identified 34 (13.1%) variants with complex genomic structures as well as tandem (193/259, 74.5%) and NAHR- mediated (6/259, 2.3%) duplications. As whole genome sequencing-based studies expand in scale and reach, a continued focus on generating high-quality, standardized duplication data will be prerequisite to addressing their associated biological mechanisms. ii Acknowledgements I thank Dr. Stephen Scherer for his leadership par excellence, his generosity, and for giving me a chance. I am grateful for his investment and the opportunities afforded me, from which I have learned and benefited. I would next thank Drs.
    [Show full text]
  • A Role for the RNA Pol II–Associated PAF Complex in AID-Induced Immune Diversification
    Article A role for the RNA pol II–associated PAF complex in AID-induced immune diversification Katharina L. Willmann,1,2 Sara Milosevic,4 Siim Pauklin,1 Kerstin-Maike Schmitz,2 Gopinath Rangam,1,2 Maria T. Simon,1 Sarah Maslen,3 Mark Skehel,3 Isabelle Robert,4 Vincent Heyer,4 Ebe Schiavo,4 Bernardo Reina-San-Martin,4 Svend K. Petersen-Mahrt1,2 1DNA Editing Laboratory, London Research Institute, South Mimms EN6 3LD, England, UK 2DNA Editing in Immunity and Epigenetics, IFOM-Fondazione Instituto FIRC di Oncologia Molecolare, Via Adamello 16, 20139 Milano, Italy 3Protein Analysis and Proteomics Laboratory, London Research Institute, South Mimms EN6 3LD, England, UK 4Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Institut National de la Santé et de la Recherche Médicale (INSERM) U964, Centre National de la Recherche Scientifique (CNRS) UMR7104, Université de Strasbourg, 67404 Illkirch, France Antibody diversification requires the DNA deaminase AID to induce DNA instability at immunoglobulin (Ig) loci upon B cell stimulation. For efficient cytosine deamination, AID requires single-stranded DNA and needs to gain access to Ig loci, with RNA pol II transcrip- tion possibly providing both aspects. To understand these mechanisms, we isolated and characterized endogenous AID-containing protein complexes from the chromatin of diversi- fying B cells. The majority of proteins associated with AID belonged to RNA polymerase II elongation and chromatin modification complexes. Besides the two core polymerase sub- units, members of the PAF complex, SUPT5H, SUPT6H, and FACT complex associated with AID. We show that AID associates with RNA polymerase-associated factor 1 (PAF1) through its N-terminal domain, that depletion of PAF complex members inhibits AID-induced immune diversification, and that the PAF complex can serve as a binding platform for AID on chromatin.
    [Show full text]
  • De Novo Variants in SUPT16H Cause Neurodevelopmental Disorders
    Novel disease loci J Med Genet: first published as 10.1136/jmedgenet-2019-106193 on 10 January 2020. Downloaded from SHORT REPORT De novo variants in SUPT16H cause neurodevelopmental disorders associated with corpus callosum abnormalities Roya Bina ,1 Dena Matalon,2 Brieana Fregeau,1 Jacqueline Joani Tarsitano,1 Ingvild Aukrust,3 Gunnar Houge,3 Renee Bend,4 Hannah Warren,4 Roger E Stevenson,4 Kyra Eva Stuurman ,5 A James Barkovich,6 Elliott H. Sherr1 1Neurology, UCSF, San Francisco, ABSTRact novo variants in the gene SUPT16H, encoding a California, USA 2 Introduction Whole- exome sequencing (WES) has subunit of the FACT (facilitates chromatin tran- P ediatrics, Stanford University, scription) complex, a histone chaperone complex Stanford, California, USA identified de novo variants in chromatin remodelling 3Department of Medical genes in patients with neurodevelopmental disorders that regulates DNA replication, transcription and Genetics, Haukeland University (NDD). We report on a novel genetic discovery in DNA repair. In our cohort, neurobehavioural chal- Hospital, Bergen, Norway lenges were prominent, including global develop- 4 chromatin remodelling in patients with NDD who also Greenwood Genetic Center, have corpus callosum (CC) anomalies. mental delay (GDD), autistic features and epilepsy. Greenwood, South Carolina, USA Objective To discover novel genes linked to both CC CC anomalies were noted in the three patients for 5Clinical Genetics, Erasmus MC, anomalies and NDD. whom brain MRIs were available for review. These Rotterdam, The Netherlands Methods Clinical WES was performed for evaluation consistent features in individuals with SUPT16H 6 Department of Radiology, of NDD, identifying five patients with de novo variants de novo variants point to a novel CC dysgenesis UCSF, San Francisco, California, disorder.
    [Show full text]
  • Datasheet: MCA3752Z Product Details
    Datasheet: MCA3752Z Description: MOUSE ANTI HUMAN SUPT16H:Preservative Free Specificity: SUPT16H Format: Preservative Free Product Type: Monoclonal Antibody Clone: 1D12 Isotype: IgG2a Quantity: 0.1 mg Product Details Applications This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit www.bio-rad-antibodies.com/protocols. Yes No Not Determined Suggested Dilution Western Blotting 0.1 - 10 ug/ml Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls. Target Species Human Species Cross Reacts with: Rat Reactivity N.B. Antibody reactivity and working conditions may vary between species. Product Form Purified IgG - liquid Preparation Purified IgG prepared by affinity chromatography on Protein A Buffer Solution Phosphate buffered saline Preservative None present Stabilisers Approx. Protein Ig concentration 0.5 mg/ml Concentrations Immunogen Recombinant protein corresponding to aa 608-716 of human SUPT16H. External Database Links UniProt: Q9Y5B9 Related reagents Entrez Gene: Page 1 of 3 11198 SUPT16H Related reagents Synonyms FACT140, FACTP140 Fusion Partners Spleen cells from immunised Balb/c mice were fused with cells from the Sp2/0 myeloma cell line. Specificity Mouse anti Human Supt16H antibody, clone 1D12 recognizes human FACT complex subunit SPT16, also known as SUPT16H, Chromatin-specific transcription elongation factor 140 kDa subunit, FACT 140 kDa subunit, FACTp140 or Facilitates chromatin transcription complex subunit SPT16.
    [Show full text]
  • Independent Study Project University of California, San Diego Abstract
    Independent Study Project University of California, San Diego Title of Project: Genomic correlates of MRI contrast enhancement in glioblastoma Author: Jeffrey M. Treiber Abstract In magnetic resonance imaging (MRI) gadolinium‐based contrast agents are administered to identify areas of blood brain barrier compromise that result in brain tumor enhancement1. The most common primary brain tumor, glioblastoma, is characterized on MRI by its heterogeneous uptake of gadolinium contrast2. Both the heterogeneity and magnitude of contrast enhancement helps clinicians to distinguish glioblastoma from other primary brain tumors3 as well as intracranial metastases4. Unfortunately, the precise molecular mechanisms which govern unique patterns of contrast enhancement remain poorly understood. Furthermore, It has been observed that glioblastoma tumors demonstrate widely different imaging characteristics and degrees of contrast enhancement. Previous studies have demonstrated relationships between contrast enhancement and overall survival as well as tumor gene expression of pathways related to cell division and hypoxia5. However, these studies were limited by small sample sizes5 and coarse qualitative metrics5‐9. Using The Cancer Imaging Archive (TCIA), a method of automated segmentation developed by our laboratory10, and matched tumor genomic data from The Cancer Genome Atlas (TCGA) this study examined the genomic correlates of quantitative measures of contrast enhancement (CE). Metrics for contrast enhancement included relative CE (rCE), CE heterogeneity, and ratio of total CE volume to the volume of the filled CE ring (CE:CEfilled). Analyses revealed relationships between rCE and inflammation, CE heterogeneity and angiogenesis, and CE:CEfilled with cell division. Age‐adjusted cox regression found a significant overall survival benefit to patients with higher CE heterogeneity (HR = 0.36, p < 0.01).
    [Show full text]
  • Omaima Nasir1
    Available online freely at www.isisn.org Bioscience Research Print ISSN: 1811-9506 Online ISSN: 2218-3973 Journal by Innovative Scientific Information & Services Network RESEARCH ARTICLE BIOSCIENCE RESEARCH, 2020 17(1): 327-346. OPEN ACCESS Exploring the Physiological and Molecular Mechanism of the Gum arabic ( Acacia senegal ) Based on Gene Expression Microarray Omaima Nasir1 1Department of Biology, Turabah University College, Taif University, Saudi Arabia. *Correspondence: [email protected], [email protected] Received 22-01-2019, Revised: 16-02-2020, Accepted: 20- 02-2020 e-Published: 01-03-2020 To investigate the differential expression of genes after Gum Arabic (Acacia senegal) administration by microarray study.The microarray was performed using colonic tissue of BALB/c s at 8 weeks of age treated with10% (w/w) GA dissolved in drinking water and control had a normal tap water for 4 days. A total 100 genes were analyzed by the pathway, gene ontology (GO) enrichment analysis, construction of protein-protein interaction (PPI) network, module analysis, construction of target genes—miRNA interaction network, target and genes-transcription factor (TF) interaction. We discuss key issues pertaining to experimental design, data preprocessing, and gene selection methods. Common types of data representation are illustrated. A total of 100 differentially miRNAs were screened and identified by microarray according to fold change the top genes which were significantly 59 genes with up-regulation and 41 genes down regulation, after Gum Arabic administration for 4 days. The data may unravel the future molecular mechanisms of Gum arabic by applying various bio-informatical approaches, we have revealed top score upregulation genes were 23, and down regulated genes with top score were 38.
    [Show full text]
  • Molecular Targeting and Enhancing Anticancer Efficacy of Oncolytic HSV-1 to Midkine Expressing Tumors
    University of Cincinnati Date: 12/20/2010 I, Arturo R Maldonado , hereby submit this original work as part of the requirements for the degree of Doctor of Philosophy in Developmental Biology. It is entitled: Molecular Targeting and Enhancing Anticancer Efficacy of Oncolytic HSV-1 to Midkine Expressing Tumors Student's name: Arturo R Maldonado This work and its defense approved by: Committee chair: Jeffrey Whitsett Committee member: Timothy Crombleholme, MD Committee member: Dan Wiginton, PhD Committee member: Rhonda Cardin, PhD Committee member: Tim Cripe 1297 Last Printed:1/11/2011 Document Of Defense Form Molecular Targeting and Enhancing Anticancer Efficacy of Oncolytic HSV-1 to Midkine Expressing Tumors A dissertation submitted to the Graduate School of the University of Cincinnati College of Medicine in partial fulfillment of the requirements for the degree of DOCTORATE OF PHILOSOPHY (PH.D.) in the Division of Molecular & Developmental Biology 2010 By Arturo Rafael Maldonado B.A., University of Miami, Coral Gables, Florida June 1993 M.D., New Jersey Medical School, Newark, New Jersey June 1999 Committee Chair: Jeffrey A. Whitsett, M.D. Advisor: Timothy M. Crombleholme, M.D. Timothy P. Cripe, M.D. Ph.D. Dan Wiginton, Ph.D. Rhonda D. Cardin, Ph.D. ABSTRACT Since 1999, cancer has surpassed heart disease as the number one cause of death in the US for people under the age of 85. Malignant Peripheral Nerve Sheath Tumor (MPNST), a common malignancy in patients with Neurofibromatosis, and colorectal cancer are midkine- producing tumors with high mortality rates. In vitro and preclinical xenograft models of MPNST were utilized in this dissertation to study the role of midkine (MDK), a tumor-specific gene over- expressed in these tumors and to test the efficacy of a MDK-transcriptionally targeted oncolytic HSV-1 (oHSV).
    [Show full text]