Protocol for measuring interorganelle contact sites in primary cells using a modified proximity ligation assay

Ilamathi, Hema Saranya; Benhammouda, Sara; Chatel-Chaix, Laurent ORCID: https://orcid.org/0000-0002-7390-8250 et Germain, Marc (2024). Protocol for measuring interorganelle contact sites in primary cells using a modified proximity ligation assay STAR Protocols , vol. 5 , nº 102915. pp. 1-15. DOI: 10.1016/j.xpro.2024.102915.

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Résumé

Interorganelle contact sites regulate lipid metabolism, organelle dynamics and positioning, as well as apoptosis and autophagy. Here, we present a proximity ligation assay (PLA) protocol for measuring the association of two organelles in fixed cells. We describe steps for primary cell culture, primary cell transfection, and the assay itself. We then detail procedures for manual and image J-based analysis of PLA foci. This protocol optimizes the use of assay products and improves the identification of PLA foci labeling actual contact sites.

Type de document:	Article
Mots-clés libres:	Cell Biology; Cell culture; Cell-based Assays; Microscopy; Molecular/Chemical Probes
Centre:	Centre INRS-Institut Armand Frappier
Date de dépôt:	27 févr. 2024 19:28
Dernière modification:	27 févr. 2024 19:28
URI:	https://espace.inrs.ca/id/eprint/14331

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