A Fast and Efficient Approach to Obtaining High-Purity Glioma Stem Cell Culture
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SOAT1 Polyclonal Antibody
PRODUCT DATA SHEET Bioworld Technology,Inc. SOAT1 polyclonal antibody Catalog: BS8121 Host: Rabbit Reactivity: Human,Mouse,Rat BackGround: Purification&Purity: SOAT1 (sterol O-acyltransferase-1), also designated The antibody was affinity-purified from rabbit antiserum ACAT1, is a homotetra-meric enzyme that catalyzes the by affinity-chromatography using epitope-specific im- formation of cholesterol esters from cholesterol and long munogen and the purity is > 95% (by SDS-PAGE). chain fatty acyl-coenzyme A (acyl-CoA). The gene en- Applications: coding human SOAT1 maps to chromosome 1 and is ex- WB 1:500 - 1:2000 pressed as a protein that localizes to the endoplasmic re- Storage&Stability: ticulum (ER) in several tissues, including liver, kidney, Store at 4°C short term. Aliquot and store at -20°C long adrenal glands and macrophages. SOAT1 is involved in term. Avoid freeze-thaw cycles. cellular cholesterol homeostasis as well as in foam cell Specificity: formation and the subsequent progression of atheroscle- SOAT1 polyclonal antibody detects endogenous levels of rosis. Several SOAT inhibitors have been developed for SOAT1 protein. the treatment of atherosclerosis. SOAT2 (sterol DATA: O-acyltransferase-2), also known as ACAT2 (acyl-CoA:cholesterol acyltransferase-2), participates in lipo-protein assembly, catalyzing cholesterol esterifica- tion in mammalian cells. SOAT2 is an integral membrane protein that localizes to the endoplasmic reticulum of human intestinal cells. SOAT2 deficiency contributes to severe mental retardation and hypotonus. Product: WesternBlot (WB) analysis of SOAT1 polyclonal antibody Rabbit IgG, 1mg/ml in PBS with 0.02% sodium azide, Note: 50% glycerol, pH7.2 For research use only, not for use in diagnostic procedure. -
ACAT) in Cholesterol Metabolism: from Its Discovery to Clinical Trials and the Genomics Era
H OH metabolites OH Review Acyl-Coenzyme A: Cholesterol Acyltransferase (ACAT) in Cholesterol Metabolism: From Its Discovery to Clinical Trials and the Genomics Era Qimin Hai and Jonathan D. Smith * Department of Cardiovascular & Metabolic Sciences, Cleveland Clinic, Cleveland, OH 44195, USA; [email protected] * Correspondence: [email protected]; Tel.: +1-216-444-2248 Abstract: The purification and cloning of the acyl-coenzyme A: cholesterol acyltransferase (ACAT) enzymes and the sterol O-acyltransferase (SOAT) genes has opened new areas of interest in cholesterol metabolism given their profound effects on foam cell biology and intestinal lipid absorption. The generation of mouse models deficient in Soat1 or Soat2 confirmed the importance of their gene products on cholesterol esterification and lipoprotein physiology. Although these studies supported clinical trials which used non-selective ACAT inhibitors, these trials did not report benefits, and one showed an increased risk. Early genetic studies have implicated common variants in both genes with human traits, including lipoprotein levels, coronary artery disease, and Alzheimer’s disease; however, modern genome-wide association studies have not replicated these associations. In contrast, the common SOAT1 variants are most reproducibly associated with testosterone levels. Keywords: cholesterol esterification; atherosclerosis; ACAT; SOAT; inhibitors; clinical trial Citation: Hai, Q.; Smith, J.D. Acyl-Coenzyme A: Cholesterol Acyltransferase (ACAT) in Cholesterol Metabolism: From Its 1. Introduction Discovery to Clinical Trials and the The acyl-coenzyme A:cholesterol acyltransferase (ACAT; EC 2.3.1.26) enzyme family Genomics Era. Metabolites 2021, 11, consists of membrane-spanning proteins, which are primarily located in the endoplasmic 543. https://doi.org/10.3390/ reticulum [1]. -
Podoplanin Regulates Mammary Stem Cell Function and Tumorigenesis by Potentiating Wnt/Β-Catenin Signaling Laura Bresson1,2,3, Marisa M
© 2018. Published by The Company of Biologists Ltd | Development (2018) 145, dev160382. doi:10.1242/dev.160382 STEM CELLS AND REGENERATION RESEARCH ARTICLE Podoplanin regulates mammary stem cell function and tumorigenesis by potentiating Wnt/β-catenin signaling Laura Bresson1,2,3, Marisa M. Faraldo1,4, Amandine Di-Cicco1, Miguel Quintanilla5, Marina A. Glukhova1,4 and Marie-Ange Deugnier1,4,* ABSTRACT K5/14), P-cadherin, smooth muscle-specific contractile proteins, Δ Stem cells (SCs) drive mammary development, giving rise postnatally and the transcription factors Np63 (an isoform of Trp63) and Slug/ to an epithelial bilayer composed of luminal and basal myoepithelial Snail2, which are essential for the maintenance of basal cell identity cells. Dysregulation of SCs is thought to be at the origin of certain breast (Yalcin-Ozuysal et al., 2010; Guo et al., 2012). The luminal cell cancers; however, the molecular identity of SCs and the factors layer is characterized by the expression of K8/18. It includes a regulating their function remain poorly defined. We identified the subset of hormone-sensing cells that express estrogen, progesterone transmembrane protein podoplanin (Pdpn) as a specific marker of the and prolactin receptors (ER, PR and PrlR, respectively) and produce basal compartment, including multipotent SCs, and found Pdpn local mediators involved in the paracrine control of basal and localized at the basal-luminal interface. Embryonic deletion of Pdpn luminal cell function (Brisken and Ataca, 2015). targeted to basal cells diminished basal and luminal SC activity and It is established that both mammary lineages, basal and luminal, affected the expression of several Wnt/β-catenin signaling components originate from a common embryonic stem cell (SC) expressing basal in basal cells. -
Role of De Novo Cholesterol Synthesis Enzymes in Cancer Jie Yang1,2, Lihua Wang1,2, Renbing Jia1,2
Journal of Cancer 2020, Vol. 11 1761 Ivyspring International Publisher Journal of Cancer 2020; 11(7): 1761-1767. doi: 10.7150/jca.38598 Review Role of de novo cholesterol synthesis enzymes in cancer Jie Yang1,2, Lihua Wang1,2, Renbing Jia1,2 1. Department of Ophthalmology, Ninth People’s Hospital of Shanghai, Shanghai Jiao Tong University School of Medicine, Shanghai, China. 2. Shanghai Key Laboratory of Orbital Diseases and Ocular Oncology, Shanghai, China. Corresponding authors: Renbing Jia, [email protected] and Lihua Wang, [email protected]. Department of Ophthalmology, Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, No. 639 Zhi Zao Ju Road, Shanghai 200011, China © The author(s). This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). See http://ivyspring.com/terms for full terms and conditions. Received: 2019.07.21; Accepted: 2019.11.30; Published: 2020.01.17 Abstract Despite extensive research in the cancer field, cancer remains one of the most prevalent diseases. There is an urgent need to identify specific targets that are safe and effective for the treatment of cancer. In recent years, cancer metabolism has come into the spotlight in cancer research. Lipid metabolism, especially cholesterol metabolism, plays a critical role in membrane synthesis as well as lipid signaling in cancer. This review focuses on the contribution of the de novo cholesterol synthesis pathway to tumorigenesis, cancer progression and metastasis. In conclusion, cholesterol metabolism could be an effective target for novel anticancer treatment. Key words: metabolic reprogramming, de novo cholesterol synthesis, cancer progress Introduction Over the past few decades, numerous published that cholesterol plays a critical role in cancer studies have focused on cancer cell metabolism and progression15-19. -
Multivariate Analysis Reveals Differentially Expressed Genes
www.nature.com/scientificreports OPEN Multivariate analysis reveals diferentially expressed genes among distinct subtypes of difuse astrocytic gliomas: diagnostic implications Nerea González‑García1,2, Ana Belén Nieto‑Librero1,2, Ana Luisa Vital3, Herminio José Tao4, María González‑Tablas2,5,6, Álvaro Otero2, Purifcación Galindo‑Villardón1,2, Alberto Orfao2,5,6 & María Dolores Tabernero2,5,6,7* Diagnosis and classifcation of gliomas mostly relies on histopathology and a few genetic markers. Here we interrogated microarray gene expression profles (GEP) of 268 difuse astrocytic gliomas—33 difuse astrocytomas (DA), 52 anaplastic astrocytomas (AA) and 183 primary glioblastoma (GBM)—based on multivariate analysis, to identify discriminatory GEP that might support precise histopathological tumor stratifcation, particularly among inconclusive cases with II–III grade diagnosed, which have diferent prognosis and treatment strategies. Microarrays based GEP was analyzed on 155 difuse astrocytic gliomas (discovery cohort) and validated in another 113 tumors (validation set) via sequential univariate analysis (pairwise comparison) for discriminatory gene selection, followed by nonnegative matrix factorization and canonical biplot for identifcation of discriminatory GEP among the distinct histological tumor subtypes. GEP data analysis identifed a set of 27 genes capable of diferentiating among distinct subtypes of gliomas that might support current histological classifcation. DA + AA showed similar molecular profles with only a few discriminatory genes -
Role of RUNX1 in Aberrant Retinal Angiogenesis Jonathan D
Page 1 of 25 Diabetes Identification of RUNX1 as a mediator of aberrant retinal angiogenesis Short Title: Role of RUNX1 in aberrant retinal angiogenesis Jonathan D. Lam,†1 Daniel J. Oh,†1 Lindsay L. Wong,1 Dhanesh Amarnani,1 Cindy Park- Windhol,1 Angie V. Sanchez,1 Jonathan Cardona-Velez,1,2 Declan McGuone,3 Anat O. Stemmer- Rachamimov,3 Dean Eliott,4 Diane R. Bielenberg,5 Tave van Zyl,4 Lishuang Shen,1 Xiaowu Gai,6 Patricia A. D’Amore*,1,7 Leo A. Kim*,1,4 Joseph F. Arboleda-Velasquez*1 Author affiliations: 1Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, 20 Staniford St., Boston, MA 02114 2Universidad Pontificia Bolivariana, Medellin, Colombia, #68- a, Cq. 1 #68305, Medellín, Antioquia, Colombia 3C.S. Kubik Laboratory for Neuropathology, Massachusetts General Hospital, 55 Fruit St., Boston, MA 02114 4Retina Service, Massachusetts Eye and Ear Infirmary, Department of Ophthalmology, Harvard Medical School, 243 Charles St., Boston, MA 02114 5Vascular Biology Program, Boston Children’s Hospital, Department of Surgery, Harvard Medical School, 300 Longwood Ave., Boston, MA 02115 6Center for Personalized Medicine, Children’s Hospital Los Angeles, Los Angeles, 4650 Sunset Blvd, Los Angeles, CA 90027, USA 7Department of Pathology, Harvard Medical School, 25 Shattuck St., Boston, MA 02115 Corresponding authors: Joseph F. Arboleda-Velasquez: [email protected] Ph: (617) 912-2517 Leo Kim: [email protected] Ph: (617) 912-2562 Patricia D’Amore: [email protected] Ph: (617) 912-2559 Fax: (617) 912-0128 20 Staniford St. Boston MA, 02114 † These authors contributed equally to this manuscript Word Count: 1905 Tables and Figures: 4 Diabetes Publish Ahead of Print, published online April 11, 2017 Diabetes Page 2 of 25 Abstract Proliferative diabetic retinopathy (PDR) is a common cause of blindness in the developed world’s working adult population, and affects those with type 1 and type 2 diabetes mellitus. -
Transcriptional Control of Lung Alveolar Type 1 Cell Development and Maintenance by NK Homeobox 2-1
Transcriptional control of lung alveolar type 1 cell development and maintenance by NK homeobox 2-1 Danielle R. Littlea,b, Kamryn N. Gerner-Mauroa, Per Flodbyc, Edward D. Crandallc, Zea Borokc, Haruhiko Akiyamad, Shioko Kimurae, Edwin J. Ostrina,f, and Jichao Chena,1 aDepartment of Pulmonary Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX 77030; bUniversity of Texas Health Graduate School of Biomedical Sciences, The University of Texas MD Anderson Cancer Center, Houston, TX 77030; cDivision of Pulmonary, Critical Care and Sleep Medicine, Department of Medicine and Hastings Center for Pulmonary Research, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033; dDepartment of Orthopedics, Kyoto University, Sakyo, 606-8507 Kyoto, Japan; eLaboratory of Metabolism, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892; and fDepartment of General Internal Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX 77030 Edited by Clifford J. Tabin, Harvard Medical School, Boston, MA, and approved August 30, 2019 (received for review April 18, 2019) The extraordinarily thin alveolar type 1 (AT1) cell constitutes nearly Hippo signaling promotes progenitor differentiation toward the the entire gas exchange surface and allows passive diffusion of AT1 cell fate (12–14). This growing list of AT1 cell regulators oxygen into the blood stream. Despite such an essential role, the highlights both the underlying complexity and the necessity to transcriptional network controlling AT1 cells remains unclear. Using distinguish direct effects on AT1 cells versus those on progenitors, cell-specific knockout mouse models, genomic profiling, and 3D imag- AT2 cells, or tissue morphology, especially in light of the classical Nkx2-1 ing, we found that NK homeobox 2-1 ( )isexpressedinAT1 observation of rapid AT1 cell-like differentiation of cultured cells and is required for the development and maintenance of AT1 AT2 cells (15). -
Supplementary Figures and Tables
SUPPLEMENTARY DATA Supplementary Figure 1. Isolation and culture of endothelial cells from surgical specimens of FVM. (A) Representative pre-surgical fundus photograph of a right eye exhibiting a FVM encroaching on the optic nerve (dashed line) causing tractional retinal detachment with blot hemorrhages throughout retina (arrow heads). (B) Magnetic beads (arrows) allow for separation and culturing of enriched cell populations from surgical specimens (scale bar = 100 μm). (C) Cultures of isolated cells stained positively for CD31 representing a successfully isolated enriched population (scale bar = 40 μm). ©2017 American Diabetes Association. Published online at http://diabetes.diabetesjournals.org/lookup/suppl/doi:10.2337/db16-1035/-/DC1 SUPPLEMENTARY DATA Supplementary Figure 2. Efficient siRNA knockdown of RUNX1 expression and function demonstrated by qRT-PCR, Western Blot, and scratch assay. (A) RUNX1 siRNA induced a 60% reduction of RUNX1 expression measured by qRT-PCR 48 hrs post-transfection whereas expression of RUNX2 and RUNX3, the two other mammalian RUNX orthologues, showed no significant changes, indicating specificity of our siRNA. Functional inhibition of Runx1 signaling was demonstrated by a 330% increase in insulin-like growth factor binding protein-3 (IGFBP3) RNA expression level, a known target of RUNX1 inhibition. Western blot demonstrated similar reduction in protein levels. (B) siRNA- 2’s effect on RUNX1 was validated by qRT-PCR and western blot, demonstrating a similar reduction in both RNA and protein. Scratch assay demonstrates functional inhibition of RUNX1 by siRNA-2. ns: not significant, * p < 0.05, *** p < 0.001 ©2017 American Diabetes Association. Published online at http://diabetes.diabetesjournals.org/lookup/suppl/doi:10.2337/db16-1035/-/DC1 SUPPLEMENTARY DATA Supplementary Table 1. -
Transcriptomic Profiles of High and Low Antibody Responders to Smallpox
Genes and Immunity (2013) 14, 277–285 & 2013 Macmillan Publishers Limited All rights reserved 1466-4879/13 www.nature.com/gene ORIGINAL ARTICLE Transcriptomic profiles of high and low antibody responders to smallpox vaccine RB Kennedy1,2, AL Oberg1,3, IG Ovsyannikova1,2, IH Haralambieva1,2, D Grill1,3 and GA Poland1,2 Despite its eradication over 30 years ago, smallpox (as well as other orthopox viruses) remains a pathogen of interest both in terms of biodefense and for its use as a vector for vaccines and immunotherapies. Here we describe the application of mRNA-Seq transcriptome profiling to understanding immune responses in smallpox vaccine recipients. Contrary to other studies examining gene expression in virally infected cell lines, we utilized a mixed population of peripheral blood mononuclear cells in order to capture the essential intercellular interactions that occur in vivo, and would otherwise be lost, using single cell lines or isolated primary cell subsets. In this mixed cell population we were able to detect expression of all annotated vaccinia genes. On the host side, a number of genes encoding cytokines, chemokines, complement factors and intracellular signaling molecules were downregulated upon viral infection, whereas genes encoding histone proteins and the interferon response were upregulated. We also identified a small number of genes that exhibited significantly different expression profiles in subjects with robust humoral immunity compared with those with weaker humoral responses. Our results provide evidence that differential gene regulation patterns may be at work in individuals with robust humoral immunity compared with those with weaker humoral immune responses. Genes and Immunity (2013) 14, 277–285; doi:10.1038/gene.2013.14; published online 18 April 2013 Keywords: Next-generation sequencing; mRNA-Seq; vaccinia virus; smallpox vaccine INTRODUCTION these 44 subjects had two samples (uninfected and vaccinia Vaccinia virus (VACV) is the immunologically cross-protective infected). -
Pdpn Podoplanin
1 Running title: Podoplanin and EMT NEW INSIGHTS INTO THE ROLE OF PODOPLANIN IN THE EPITHELIAL TO MESENCHYMAL TRANSITION Jaime Renart*, Patricia Carrasco-Ramírez, Beatriz Fernández-Muñoz1, Ester Martín- Villar, Lucía Montero, María M. Yurrita and Miguel Quintanilla. Instituto de Investigaciones Biomédicas Alberto Sols, CSIC-UAM *Corresponding author at: Instituto de Investigaciones Biomédicas Alberto Sols, CSIC- UAM. Arturo Duperier 4. 28029-Madrid. Spain. Tel: +34915854439. Fax: +34915854401. E-mail address: [email protected] 1Present address: Laboratorio Andaluz de Reprogramación Celular, Parque Tecnológico y Científico Cartuja 93. 41092-Sevilla, Spain. 2 CONTENTS 1. Introduction 2. Podoplanin 2.1. Protein structure 2.1.1. The ectodomain 2.1.2. The transmembrane domain 2.1.3. The cytoplasmic domain 2.2. Gene structure 2.3. Expression of podoplanin 2.3.1. Tissue distribution 2.3.2. Transcriptional regulation 2.3.3. Post-translational regulation 2.3.3.1. miRNAs 2.3.3.2. Glycosylation 2.3.3.3. Proteolytic processing 2.3.3.4. Phosphorylation 2.4. Podoplanin partners 2.4.1. CLEC-2 2.4.2. Tetraspanin CD9 2.4.3. Galectin 8 2.4.4. Heat shock protein A9 2.4.5. CD44 2.4.6. ERM proteins 3 2.4.7. Others 2.5. Signaling and molecular mechanisms 2.6. Podoplanin in development 2.7. Podoplanin in cancer 2.7.1. Expression in tumors 2.7.2. Role in migration, invasion and progression 2.7.3. Presence in tumor stroma 3. Epithelial to mesenchymal transition 3.1. Molecular mechanisms of EMT 3.1.1. Properties of the core Transcription factors 3.1.2. -
Mouse Embryonic Fibroblasts Exhibit Extensive Developmental and Phenotypic Diversity
Mouse embryonic fibroblasts exhibit extensive developmental and phenotypic diversity Prabhat K. Singhala,b, Slim Sassia,c, Lan Lana,b, Patrick Aud, Stefan C. Halvorsena, Dai Fukumurad, Rakesh K. Jaind,1, and Brian Seeda,b,1 aCenter for Computational and Integrative Biology, Massachusetts General Hospital, Boston, MA 02114; bDepartment of Genetics, Harvard Medical School, Boston, MA 02115; cDepartment of Orthopedics, Harvard Medical School, Boston, MA 02115; and dSteele Laboratory of Tumor Biology, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114 Contributed by Rakesh K. Jain, November 16, 2015 (sent for review August 22, 2015; reviewed by Patricia A. D’Amore, Massimo Pinzani, and David Tuveson) Analysis of embryonic fibroblasts from GFP reporter mice indicates that light or electron microscopy, but exhibit variable GFP expression. the fibroblast cell type harbors a large collection of developmentally Populations from earlier stages, days E12.5 and E14.5, exhibit lower and phenotypically heterogeneous subtypes. Some of these cells frequencies of GFP-positive cells compared with cultures prepared exhibit multipotency, whereas others do not. Multiparameter flow from day E16.5 and E18.5 embryos (Fig. 1A). GFP-positive cells cytometry analysis shows that a large number of distinct populations typically replicate more rapidly than GFP-negative cells under stan- of fibroblast-like cells can be found in cultures initiated from different dard culture conditions, leading to an increase in their prevalence embryonic organs, and cells sorted according to their surface pheno- with increasing culture passage (Fig. 1B). type typically retain their characteristics on continued propagation in Following fluorescence-activated cell sorting of newly initiated culture. -
Molecular Taxonomy of Human Ocular Outflow Tissues Defined by Single-Cell Transcriptomics
Molecular taxonomy of human ocular outflow tissues defined by single-cell transcriptomics Gaurang Patela,1, Wen Furya,1, Hua Yanga, Maria Gomez-Caraballob, Yu Baia, Tao Yanga, Christina Adlera, Yi Weia, Min Nia, Heather Schmittb, Ying Hua, George Yancopoulosa, W. Daniel Stamerb,2, and Carmelo Romanoa,2 aRegeneron Pharmaceuticals, Inc., Tarrytown, NY 10591; and bDepartment of Ophthalmology, Duke University, Durham, NC 27710 Edited by Joel S. Schuman, New York University, New York, NY, and accepted by Editorial Board Member Jeremy Nathans April 16, 2020 (received for review January 31, 2020) The conventional outflow pathway is a complex tissue responsible refs. 7 and 8). Cells that populate the TM are all of neural crest in for maintaining intraocular pressure (IOP) homeostasis. The co- origin (13), displaying different morphologies depending upon their ordinated effort of multiple cells with differing responsibilities tissue location. Thus, TM cells in uveal/corneoscleral meshwork ensures healthy outflow function and IOP maintenance. Dysfunc- display endothelial and macrophage properties, maintaining patent tion of one or more resident cell types results in ocular hyperten- flow passageways by secreting antithrombotic molecules (14), sion and risk for glaucoma, a leading cause of blindness. In this phagocytosing cellular debris, neutralizing reactive oxygen species study, single-cell RNA sequencing was performed to generate a (15–17), and mediating immune function (18, 19), while the cells in comprehensive cell atlas of human conventional outflow tissues. the JCT region display fibroblast- and smooth muscle-like proper- We obtained expression profiles of 17,757 genes from 8,758 cells ties, playing an important role in the generation and control of from eight eyes of human donors representing the outflow cell outflow resistance (7).