Densitometry done twice. Trend is the same. p62 and ATG7 are both decreased in the treated cells vs mock.
The decrease in LC3-I expression suggests the conversion of LC-3, and the parallel decrease of LC3-II implies the degradation of autophagolysosome by protease. So that it is likely that the autophagic process is quite rapid in your experimental cells.
Thank you so much. What is your suggestion to check the speeding of autophagosome degradation by treatment?
Another possibility might be degradation of total LC3 and reduction in autophagy. But it can only be confirmed by using BafilomycinA or 3-MA.
I think the decrease in both LC3I and LC3II signifies effect on LC3B expression rather than on LC3 conversion which is what classically thought and explained as LC3I is decreased and LC3 II is increased during autophagy process. The continuing approach could be pretreatment with bafilomycin as suggested previously. Another point i would like to add is during western blotting the antibody used may not have sensitivity as such to see corresponding decrease in Lc3I and increase in Lc3II for autophagy increment. So, i suggest rather than looking at both LC3 I and LC3II, compare based on LC3II, perform additional experiments using bafilomycin,p62 stuffs, and other techniques for further confirmation
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