Fig. 5
miR-30c-1-3p inhibits autophagy by targeting ATG4B and ATG9B during H37Rv infection. (A) Western Blot demonstrated that miR-30c-1-3p mimics intervention reduced H37Rv-induced increase in LC3-II, (B) ATG4B and ATG9B in macrophages. The LC3, ATG4B and ATG9B bands were quantified relative to β-actin. Since A and B are the same experiment with the same batch of cells, the same β-actin blot was used. (C) Confocal microscopy detected LC3 puncta in macrophages infected with H37Rv for 2 h. The number of LC3 puncta was significantly reduced in miR-30c-1-3p mimics transfected macrophages. Scale bars: 2 μm. (D) The inhibition of autophagy by miR-30c-1-3p was confirmed by transmission electron microscopy. In macrophages infected with H37Rv for 2 h, an increase in the number of autophagolysosomes was observed, while a decrease in the number of autophagolysosomes was observed in macrophages transfected with miR-30c-1-3p mimics. Green and red arrows represent suspected autophagosomes and autophagolysosomes respectively, and the number of autophagolysosomes was counted for each cell cross-section. Scale bars: 5 μm. All data above represent the means ± SD of at least three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not statistically different.
