title: Analysis of Nucleocytoplasmic Protein Shuttling by Imaging Flow Cytometry.
abstract: Many intracellular signal transduction events involve the reversible shuttling of proteins between the cytoplasm and the nucleus. Study of these processes requires imaging information on the protein localization in a given cell and a large number of measurements to obtain sufficient statistics on the protein localization in the whole population. The protocol describes method for quantitative imaging flow cytometry analysis of intracellular distribution of NF-kappaB in ARPE-19 cells stained with specific fluorochrome-conjugated antibodies. The described technique alone or in combination with standard flow cytometry methods can be applied to study any protein undergoing translocation from cytoplasm into the nucleus in a variety of cell lines as well as in heterogeneous primary cell populations.
authors.affiliation: Department of Biomedicine, University of Basel and University Hospital Basel, CH-4031, Basel, Switzerland. elizaveta.fasler@insel.ch.|Department of Pediatric Surgery and Department of Clinical Research, Inselspital University Hospital, University of Bern, Bern, CH-3010, Switzerland. elizaveta.fasler@insel.ch.
authors.forename: Elizaveta
authors.initials: E
authors.lastname: Fasler-Kan
authors.affiliation: Nazarbayev University, Astana, Kazakhstan.|Cellular and Molecular Medicine Program, Boston Children's Hospital, Boston, MA, USA.
authors.forename: Yeldar
authors.initials: Y
authors.lastname: Baiken
authors.affiliation: Department of Cell Biology and Histology, M.V. Lomonosov Moscow State University, Moscow, 119992, Russia.|A.N. Belozersky Institute of Physico-Chemical Biology, M.V. Lomonosov Moscow State University, Moscow, 119992, Russia.
authors.forename: Ivan A
authors.initials: IA
authors.lastname: Vorobjev
authors.affiliation: Nazarbayev University, Astana, Kazakhstan.|Department of Pediatrics, Harvard Medical School, Boston, MA, USA.
authors.forename: Natasha S
authors.initials: NS
authors.lastname: Barteneva
mesh_terms: D021581:Active Transport, Cell Nucleus; D002460:Cell Line; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D019044:Image Cytometry / Q000379:methods*; D016328:NF-kappa B / Q000378:metabolism
chemical_list: D016328:NF-kappa B
keywords: ARPE-19 cell line; Flow cytometry; Imaging cytometry; NF-kappaB; Nucleocytoplasmic shuttling; Nucleocytoplasmic translocation; Retinal pigment cells