Figure 4.

6-Sho enhances the TRAIL-initiated apoptotic pathway by attenuating autophagy flux. Huh7 cells were pretreated with CQ (10 mM) for 1 h prior to exposure to 6-sho (20 µM) for 18 h and TRAIL (200 ng/ml) for 2 h. (A) LC3-II and p62 levels assessed by western blotting using β-actin as control. (B) Cells stained with p62 antibody (green) and DAPI nuclear stain (blue) evaluated using fluorescence microscopy (magnification, ×400). (C) Cas3 and cas8 levels assessed by western blot assays using β-actin as control. 6-Sho, 6-shogaol; TRAIL, tumor necrosis factor-related apoptosis-inducing ligand; CQ, chloroquine; p62, tumor-suppressor protein 62; LC3, microtubule-associated proteins 1A/1B light chain 3B; cas, caspase.