Fig. 1.

miR-26a overexpression promotes autophagy. a Cells were transfected with miR-26a or scramble miRNA (miR-NC) for 24 h. Then, cells were treated with HBSS for 4 h or 125 nM rapamycin for 24 h. The expression of p62 and LC3 was detected by immunoblotting. b SK-Hep-1 cells were transfected with miR-26a inhibitor (miR-26aI) or negative control for 24 h. Then, cells were treated with HBSS for 4 h or 125 nM rapamycin for 24 h. The expression of p62 was detected by immunoblotting. The relative quantity of p62 in a and b was calculated by ImageJ densitometric analysis and normalized using GAPDH. c Representative TEM images of SK-Hep-1 cells transfected with miR-26a for 48 h as indicated. Bar 1 μm. Arrows denote autolysosomes. d Quantification of autolysosomes (denoted by white arrows) per cell (n = 30). e Percentage of red or yellow puncta-positive cells was quantified by automated image acquisition and analysis using a threshold of more than five dots per cell. f SK-Hep-1 RLuc–LC3^WT and RLuc–LC3^G120A cells were reverse-transfected with miRNAs and siRNAs or treated with 125 nM rapamycin. Luciferase activity was measured at 24 and 48 h after transfection or rapamycin treatment. Results shown are the mean ± SD of at least three independent experiments. *P < 0.05; ** P < 0.01