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Methods Mol Biol. 2017;1557:111-115. doi: 10.1007/978-1-4939-6780-3_11.

Organelle Separation and Cell Signaling.

Author information

1
INSERM, UMR_S 1197, Hôpital Paul Brousse, 14 Avenue Paul Vaillant Couturier, 94807, Villejuif Cedex, France.
2
Université Paris-Saclay, Paris-Saclay, France.
3
INSERM, UMR_S 1197, Hôpital Paul Brousse, 14 Avenue Paul Vaillant Couturier, 94807, Villejuif Cedex, France. damien.arnoult@inserm.fr.
4
Université Paris-Saclay, Paris-Saclay, France. damien.arnoult@inserm.fr.

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.

KEYWORDS:

Cell fractionation; Differential centrifugation; Immunoblotting; Signaling; Ubiquitination

PMID:
28078587
DOI:
10.1007/978-1-4939-6780-3_11
[Indexed for MEDLINE]

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