title: Organelle Separation and Cell Signaling.
abstract: Recent findings indicate that some signaling hubs coalesce at the surfaces of organelles through the accumulation of ubiquitylated components required for the signal transduction. For instance, ubiquitylated components of the NF-κB pathway accumulated at the endoplasmic reticulum while ubiquitylated components of the IRF3 pathway are found at the Golgi apparatus. Here we describe simple methods to observe and assess these ubiquitylated components by immunoblotting using differential centrifugation and in vitro assays.
authors.affiliation: INSERM, UMR_S 1197, Hôpital Paul Brousse, 14 Avenue Paul Vaillant Couturier, 94807, Villejuif Cedex, France.|Université Paris-Saclay, Paris-Saclay, France.
authors.forename: Leandro Silva
authors.initials: LS
authors.lastname: Da Costa
authors.affiliation: INSERM, UMR_S 1197, Hôpital Paul Brousse, 14 Avenue Paul Vaillant Couturier, 94807, Villejuif Cedex, France. damien.arnoult@inserm.fr.|Université Paris-Saclay, Paris-Saclay, France. damien.arnoult@inserm.fr.
authors.forename: Damien
authors.initials: D
authors.lastname: Arnoult
mesh_terms: D015153:Blotting, Western; D002458:Cell Fractionation / Q000379:methods; D002499:Centrifugation, Density Gradient / Q000379:methods; D050838:Interferon Regulatory Factor-3 / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D015388:Organelles / Q000378:metabolism*; D010766:Phosphorylation; D015398:Signal Transduction; D054875:Ubiquitination
chemical_list: D050838:Interferon Regulatory Factor-3; D016328:NF-kappa B
keywords: Cell fractionation; Differential centrifugation; Immunoblotting; Signaling; Ubiquitination