Fig. 3

Inhibitory effect of PG on autophagy is mediated by SNAREs complex. A TFK-1 cells transfected with mRFP-GFP-LC3 were treated with DMSO (0.1%), PG (600 nM), CQ (10 µM), Rap (600 nM), and PG (600 nM) + CQ (10 µM), respectively, and the changes in green and red fluorescence were observed. Scale bar: 10 mm. B Western blot was used to detect the expression levels of SNARE proteins (STX17, SNAP29, and VAMP8) after treating CCA cells for 24 h with different concentrations of PG. C SNAP29 and STX17 were overexpressed in CCA cells, respectively. After the addition of PG or DMSO, the expression levels of LC3B-II/I and P62 autophagy proteins were detected by western blot analysis. D After transfecting TFK-1 cells with mRFP-GFP-LC3 for 24 h, autophagy flux was observed using a confocal microscope. E STX17 and SNAP29 were overexpressed in HuCCT-1 and TFK-1. After PG or DMSO treatment for 24 h, the changes in autolysosomes in CCA cells were observed by TEM. N, nuclear; arrows, autolysosomes; All data are representative of three independent experiments. *P < 0.05