Dear all,
In an in vitro experiment of autophagic flux assay by western blot using chloroquine, is it correct to use the LC3-II/LC3-I ratio as a manner to represent the effects of a treatment or should I consider just the results of LC3-II?
Thank you very much in advance.
Dear Machado,
Before analyzing Autophagy flux, I suggest first you look autophagy markers such as LC3II, p62 levels. There by you got some idea about autophagy induction. Then go for autophagy flux in presence of activators/inhibitors.
All the best
Dr.Nagesh
Dear Reddy,
Thank you very much for your message. I have looked already to LC3II, and in fact the ratio of LC3II/LC3I was increased. Then I decided to perfom the autophagic flux assay just to better confirm that previous effect. However, I have obtained results quite difficult to interpret.
The isolated expression of LC3II has no value in itself, it must be reported as a ratio to that of actin (LC3II / actin) or that of LC3I (LC3II / LC3I).
Ref: Klionsky DJ, Abdalla FC, Abeliovich H, et al. Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy 2012;8:445-544.
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