I want to detect the kinetics of the activation of IRE1, PERK and ATF6 at single cell level. Ideally cells expressing reporter constructs of equal efficiency to be able to detect temporal activation each of the 3 arm of the UPR.
Dear Dr. Samali
It is a very interesting question. I believe if you infect the cells with reporter construct and use laser scanning cytometry (LSC)which is equipped with chamber for growing the cells you can do this assay. LSC is a strong tool and has lots of ability which is the combination of FACS and an epi fluorescence microscope. I would be happy to discuss in more detail.
Please review the following article:
PMID: 16719355 [PubMed - indexed for MEDLINE] PMCID: PMC3892962
Also you can see my BBA-MCR paper (2010) and CDDis paper (2012) that we have use LSC to detect apoptosis.
Regards
Saeid
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