Protocol for measuring interorganelle contact sites in primary cells using a modified proximity ligation assay.
STAR Protoc
; 5(1): 102915, 2024 Mar 15.
Article
in En
| MEDLINE
| ID: mdl-38393951
ABSTRACT
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. For complete details on the use and execution of this protocol, please refer to Ilamathi et al. (2023).1.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Autophagy
/
Apoptosis
Language:
En
Journal:
STAR Protoc
/
STAR protocols
Year:
2024
Document type:
Article
Country of publication: