Should it be more valuable to observe an increased ratio of LC3-I to Actin rather than see decreasing ratio of LC3-II to Actin in the presence of drugs inhibit autophagy?
The problem is. that antibodies against LC3 recognize the LC3-II form more reliably than the LC3-I form, such that autophagy is measured as LC3-II/housekeeper. The LC3-I signal also depends on how many times the extracts were thawed.
For more info, see: Daniel J. Klionsky, et al., Autophagy 2012 Vol. 8:4, 445–544; Guidelines for the use and interpretation of assays for monitoring autophagy.