Fig. 6: PIASy induces autophagpy flux in Huh7 cells.

In the presence or absence of an autophagosome–lysosome fusion inhibitor, chloroquine (CQ, 25 uM), Huh7 cells (a, b), or Huh7 cells expressing GFP-LC3 (c) were transfected with pCMV-myc-PIASy/empty vector (a, c) or shRNA-PIASy/control-scrambled shRNA (b, c) for 96 h. a A representative western blot image shows the protein levels of PIASy, LC3B-I, LC3B-II, and p62 in PIASy-overexpressed or control cells. The relative ratios of PIASy/β-actin, LC3B-II/β-actin, LC3B-II/LC3B-I, and p62/β-actin were calculated and shown as the fold of control (without CQ treatment and with empty vector transfection, which was defined as 1). b A representative western blot image shows the protein levels of PIASy, LC3B-I, LC3B-II, and p62 in PIASy-down-expressed or control cells. The relative ratios of PIASy/β-actin, LC3B-II/β-actin, LC3B-II/LC3B-I, and p62/β-actin were calculated and shown as the fold of control (without CQ treatment and with scrambled shRNA transfection, which was defined as 1). c GFP-LC3-positive dots per cell were determined by fluorescence microscope (30 cells were counted per experiment) in PIASy-overexpressed, PIASy-down-expressed, or control cells. Scale bar: 50 µm. The data are the mean ± SD of the results of three independent experiments. *p < 0.05, **p < 0.01