Fig. 3

MiR-214 regulated autophagy at both basal and 4-OHT/FUL induced level. a MCF7 cells were treated with 5 μM 4-OHT or 1 μM FUL for 48 h. Western blotting was performed to analyze LC3 and Beclin-1. Bar graphs indicated relative levels of LC3-II and Beclin-1 normalized to β-actin. **P < 0.01 vs. vehicle control. b The expression of miR-214 was analyzed by RT-qPCR. **P < 0.01 vs. vehicle control. c and d) MCF7 cells were transfected with 100 nM miR-214 mimics (214 m) or inhibitors (214i) for 24 h, then were treated with or without 5 μM 4-OHT or 1 μM FUL for 48 h. Western blotting was performed to analyze LC3-II normalized to β-actin. RT-qPCR was performed to analyze the level of miR-214 (c). *P < 0.05, **P < 0.01 vs. vehicle control, ^## P < 0.01 vs. negative control (NC). e MCF7 cells were co-transfected with GFP-LC3 and 100 nM miR-214 mimics or inhibitors for 48 h in the presence or absence of 5 mM 3-MA. Cells were imaged under a confocal microscopy (scale bar = 10 μm). Numbers of GFP-LC3 puncta per cell were counted (*P < 0.05 **P < 0.01 vs. negative control, ^## P < 0.01 vs. transfection of miR-214 inhibitors (214i), n = 10). Data represent mean ± SD of three experiments