FIGURE 7.

Gallic acid inhibits autophagy and induces cell death in SW1088 cells. (A) SW1088 cells were transfected with vector or c‐RAB13, and then treated with or without gallic acid. The expression and location of LC3 were detected by immunofluorescence. Scale bar = 20 µm. (B) After SW1088 cells were transfected with si‐RAB13 and c‐RAB13 for 24 h, after co‐incubation with gallic acid (1 mM) or 3‐MA (10 nM). Then, SW1088 cell was transfected with GFP/mRFP‐LC3 plasmid, representative images and quantitative analysis of LC3 puncta were shown. Scale bar = 20 µm. (C) SW1088 cells were transfected with vector or c‐RAB13, and then treated with or without Gallic acid for 24 h. The expression of LC3 was detected by Western blot analysis. All data were representative of at least three independent experiments. Merge compared with the NC group, # p < 0.05, ## p < 0.01, ### p < 0.001, ns, no significance; RFP compared with the NC group, *p < 0.05, ** p < 0.01, *** p < 0.001, ns, no significance. (D) SW1088 cells were treated with different concentrations of gallic acid for 24 h. The expression of RAB13 was detected by Western blot analysis. All data were representative of at least three independent experiments. GA‐L, Gallic acid 1 mM; GA‐M, Gallic acid 2 mM; GA‐H, Gallic acid 3 mM; ns, no significance; P*<0.05, **p < 0.01, ***p < 0.001. (E) SW1088 cells were treated with different concentrations of gallic acid for 24 h, and the apoptosis ratios were determined by flow cytometry analysis of Annexin‐V/PI double staining. GA‐L, Gallic acid 1 mM; GA‐L, Gallic acid 2 mM; GA‐L, Gallic acid 3 mM. (F) SW1088 cells were transfected with vector or c‐RAB13, and then treated with or without Gallic acid for 24 h, and the apoptosis ratios were determined by flow cytometry analysis of Annexin‐V/PI double staining