since I have observed that in other cells an increase in LC3-II occurs when uses EBSS or other stimuli for induces autophagy.
Hello Rayén,
In my opinion, the mere amount of LC3-II can be a missdirecting marker to look at in order to interpret autophagy induction. Different cell lines have different rates of autophagic flux, therefore LC3-II could be autolysosomally degraded already.
I suggest to do a kinetic of the autophagic flux starting at 0.5 h until 6 h or more and additionally add controls of EBSS + bafilomycin A1 (a lysosomal inhibitor, which prevents autolysosomal degradation of LC3-II).
As far as I checked, H4 cells seem to show a decreased amount of LC3-II upon EBSS already after 4 hours (e.g. Nyfeler et al., 2011 DOI: 10.1128/MCB.05430-11).
Kind regards,
Jana
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