Autophagy in anti-apoptotic effect of augmenter of liver regeneration in HepG2 cells

Figure 2 Augmenter of liver regeneration increases autophagic activity in HepG2 cells. HepG2 cells were transfected with an augmenter of liver regeneration (ALR)-expressing plasmid or a control vector and cultured in Dulbecco’s modified Eagle’s medium for 6 h, followed by treatment with serum-free medium. Autophagy was observed at 24 h after serum deprivation. A: Total cell lysates were subjected to Western blot analysis with anti-LC3, anti-p62 and anti-β-actin antibodies. Densitometry was performed using ImageJ software to analyze the expression of LC3-II; B: GFP-LC3 puncta were determined by counting a total of more than 30 cells in triplicate samples; C: Digital images were obtained using an electron microscope (H-7650, Japan). The numbers of typical autophagosomes and autolysosomes in each cell section were counted randomly for more than 30 cells; D: The expression of ATG5, ATG7 and Beclin-1 mRNA was determined by qPCR. Digital data are presented as the mean ± SD from at least three independent experiments. ^aP < 0.05 ALR (-) vs ALR (+).