Figure 4
Blockage of endogenous miR-20a promotes autophagy and lysosome activity. (a) Representative GFP-LC3 images in MDA-MB-231 cells expressing LNA-NC or LNA-20a. Scale bar, 10 μm. GFP-LC3 dots per cell was shown as means±s.d. (*P<0.05). (b) MDA-MB-231 and MCF7 cells transfected with LNA-NC or LNA-20a were cultured in normal medium or EBSS for 4 h. Relative ratios of LC3-II versus GAPDH were assessed by western blotting and Image J software. (c) Immunoblotting analysis of OPTN and ACTB in MDA-MB-231 and MCF7 cells transfected with LNA-NC or LNA-20a. (d) Quantitative analysis of autophagosomes (yellow dots; mCherry+/GFP+) and autolysosomes (red dots; mCherry+/GFP−) in MCF7 cells transfected with mCherry-GFP-LC3 and LNA-20a or LNA-NC (*P<0.05, **P<0.01). (e) Representative images of DQ-BSA in MCF7 cells overexpressing LNA-NC or LNA-20a that subjected with or without nutrient starvation for 4 h. (f) Quantification of DQ Red BSA fluorescence in MDA-MB-231 or MCF7 cells (**P<0.01).
