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CD95 pp 111-115 | Cite as

Organelle Separation and Cell Signaling

  • Leandro Silva Da Costa
  • Damien Arnoult
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1557)

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.

Key words

Differential centrifugation Cell fractionation Ubiquitination Signaling Immunoblotting 

Notes

Acknowledgment

This work was supported by grants from Fondation ARC (Association pour La Recherche contre le Cancer) and the ANRS (Agence Nationale pour la Recherche sur le SIDA).

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.INSERM, UMR_S 1197Hôpital Paul BrousseVillejuif CedexFrance
  2. 2.Université Paris-SaclayParis-SaclayFrance

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