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Quantification of eIF2α Phosphorylation Associated with Mitotic Catastrophe by Immunofluorescence Microscopy
Juliette Humeau1, Lucillia Bezu2,1, Oliver Kepp2,1
1Centre de Recherche des Cordeliers, Université de Paris, Sorbonne Université, Inserm U1138, Institut Universitaire de France, Paris, France.
Methods in Molecular Biology (Clifton, N.J.)
|March 31, 2021
Summary
Mitotic catastrophe triggers cell death or senescence via errors in mitosis. This study presents a high-throughput method to quantify eukaryotic translation initiation factor 2 alpha (eIF2α) phosphorylation, a key stress response marker.
Area of Science:
- Cell Biology
- Molecular Biology
- Cancer Research
Background:
- Mitotic catastrophe is a tumor-suppressive mechanism initiated by errors during cell division.
- Eukaryotic cells activate adaptive signaling pathways to manage cellular stress.
- Phosphorylation of eukaryotic translation initiation factor 2 alpha (eIF2α) at serine 51 is a conserved stress response.
Purpose of the Study:
- To detail a method for quantifying eIF2α phosphorylation.
- To capture the integrated stress response in the context of mitotic catastrophe.
Main Methods:
- High-throughput immunofluorescence microscopy.
- Quantification of eIF2α phosphorylation at serine 51.
Main Results:
- The described method effectively quantifies eIF2α phosphorylation.
- This technique captures the integrated stress response induced by mitotic catastrophe agents.
Conclusions:
- A novel high-throughput method allows for the quantification of eIF2α phosphorylation.
- This method aids in understanding the integrated stress response during mitotic catastrophe.
