Figure 1. Overexpression of MIR181A resulted in decreased autophagic activity in MCF-7 cells. (A) MIR181A blocked starvation-induced GFP-LC3 dot formation in MCF-7 cells. Cells were cotransfected with MIR181A or control construct (MIR-CNT) together with GFP-LC3 plasmid and autophagy was assessed under no starvation or starvation (2 h) conditions. White arrows indicate clusters of the GFP-LC3 dots in cells. (B) Quantitative analysis of the experiments in (A). MIR181A overexpression, but not control (MIR-CNT) overexpression, blocked starvation-induced autophagy (mean ± SD of independent experiments, n = 3, **p < 0.01. N.S., not significant). (C) MIR181A decreased starvation-induced conversion of LC3-I to LC3-II in MCF-7 cells. Immunoblot results of extracts from nonstarved (STV-) or starved (STV+) cells (n = 3). LC3-II/LC3-I densitometric ratios are marked. ACTB was used as a loading control. (D) MIR181A blocked starvation induced SQSTM1 degradation in MCF-7 cells (n = 3). ACTB was used as a loading control. SQSTM1/ACTB densitometric ratios were marked. (E) MIR181A blocked rapamycin-induced GFP-LC3 dot formation. Cells were cotransfected with GFP-LC3 plasmid and MIR181A or MIR-CNT, treated with DMSO (carrier) or rapamycin (2.5 µM, 24 h). (F) Quantitative analysis of the experiments in (E) (mean ± SD of independent experiments, n = 4, *p < 0.05. N.S., not significant). (G) MIR181A decreased LC3-I to LC3-II conversion stimulated by rapamycin (RAP) in MCF-7 cells (n = 2). LC3-II/LC3-I ratios are marked. (H) MIR181A blocked rapamycin-induced SQSTM1 degradation in MCF-7 cells (n = 2).