Thp-1cell treated with X drug= LC3II increase and p62 increase
Thp-1cell treated with X drug and bafilomycin =LC3II increase and p62 slightly decrease
Drug X is expected to promote the autophagic flux with increased LC-3 II and down-regulation of p62/SQSTM1.
Given that bafilomycin A1 blocks the stable autophagy flux, it seems reasonable that both LC-3 II and p62/SQSTM1 increases.
I agree with the previous comments, but the literature has shown that the LC3-II / LC3-I ratio has not been considered a good parameter for the interpretation of the autophagy flow. In my previous experience, I obtained results (without the use of drugs that inhibit or activate autophagy) in which there was an increase in LC3-II concomitant with an increase in p62. My choice to better interpret my data was to evaluate other proteins from the different stages of the autophagy flow, such as beclin1, LAMP2 and CHMP2B. As well as immunofluorescence analyzes and transmission electron microscopy.
I suggest the reading of Klionsky DJ, Abdelmohsen K, Abe A, et al. Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition) [published correction appears in Autophagy. 2016;12(2):443. Autophagy. 2016;12(1):1-222. doi:10.1080/15548627.2015.1100356, to better understand your data.
Good luck with your analysis.
I agree with the previous comments, but the literature has shown that the LC3-II / LC3-I ratio has not been considered a good parameter for the interpretation of the autophagy flow. In my previous experience, I obtained results (without the use of drugs that inhibit or activate autophagy) in which there was an increase in LC3-II concomitant with an increase in p62. My choice to better interpret my data was to evaluate other proteins from the different stages of the autophagy flow, such as beclin1, LAMP2 and CHMP2B. As well as immunofluorescence analyzes and transmission electron microscopy.
I suggest the reading of Klionsky DJ, Abdelmohsen K, Abe A, et al. Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition) [published correction appears in Autophagy. 2016;12(2):443. Autophagy. 2016;12(1):1-222. doi:10.1080/15548627.2015.1100356, to better understand your data.
Good luck with your analysis.
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