Hello all,
I treated with a type of adjuvant I am interested in with DC 2.4 cell lines (DC-like murine cells) for 3, 6, 12, and 24 hrs interval. I then lysed the whole cell extract and did WB against LC3 and p62. LC3-II band for some reason is very weak. Even with rapamycin (the end lane) is kind of weak, but LC-II band is visible compared to control (-). I tried 40 ug/well.
So, at 24 hours, LC 3 I and II bands almost disappear. Well, you can see more of LC3-II, but what does this mean? Also, when you see more of LC3-II, p62 is also increased. Does that mean autophagy is "stuck" rather than "ongoing?" If so, what does the stuck autophagy mean?
Your help is much appreciated!
Hi Monica,
i might be missing out but I didn't see the interpretation of LC3-1 "disappearing"
- Dear LEE,
- Autophagy is a dynamic phenomenon. your observation will indicate the autophagy flux where there is an increase in the number of autophagosomes degraded there by decrease in LC3 II levels. In our experiments, co localization of LC3 and p62 were observed in IFA.
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