title: Analyzing the Impact of Pan- and Class-Specific HDACi on Differentiation-Associated Factors.
abstract: The differentiation of hematopoietic stem cells into mature blood cells is a highly ordered process and dysregulation of this process can lead to leukemogenesis. Agents that are used to cure acute promyelocytic leukemia (APL) can induce differentiation and/or apoptosis. Here, we describe how effects of all-trans retinoic acid (ATRA) and histone deacetylase inhibitors (HDACi) on APL cell differentiation can be evaluated by immunoblotting and by flow cytometry. We show how the levels of differentiation-associated transcription factors of the CCAAT enhancer binding protein (C/EBP) family can be determined by Western blot and we explain how the cell surface expression of the leukocyte surface antigen CD11b can be measured by flow cytometry.
authors.affiliation: Institute for Biochemistry and Biophysics, Centre for Molecular Biomedicine (CMB), Friedrich-Schiller-Universität Jena, Hans-Knöll-Str. 2, Jena, 07745, Germany.|Integrated Research and Treatment Center, Center for Sepsis Control and Care (CSCC), Jena University Hospital, Erlanger Allee 101, Jena, 07747, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Noack
authors.affiliation: Institut für Toxikologie, Universitätsmedizin Mainz, Obere Zahlbacher Str. 67, Mainz, 55131, Germany. okraemer@uni-mainz.de.
authors.forename: Oliver H
authors.initials: OH
authors.lastname: Krämer
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D001549:Benzamides / Q000494:pharmacology; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D022762:CCAAT-Enhancer-Binding Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D039481:CD11b Antigen / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D005434:Flow Cytometry; D015973:Gene Expression Regulation, Leukemic; D006412:Hematopoietic Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D015151:Immunoblotting; D007211:Indoles / Q000494:pharmacology; D000077767:Panobinostat; D011725:Pyridines / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D001549:Benzamides; D022782:CCAAT-Enhancer-Binding Protein-beta; D022762:CCAAT-Enhancer-Binding Proteins; D039481:CD11b Antigen; C545012:CEBPB protein, human; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D007211:Indoles; D011725:Pyridines; C070790:CEBPE protein, human; C118739:entinostat; D014212:Tretinoin; D000077767:Panobinostat
keywords: APL; C/EBPβ; C/EBPε; CD11b; Flow cytometry; HDACi; Western blot