Figure 8.

Cryptotanshinone (CPT) stimulates autophagy in Huh7 and MHCC97-H cells. (A) Western blot analysis of LC3-I, LC3-II, p62/SQSTM1, Beclin1, and ATG5 in human hepatocellular carcinoma (HCC) cells treated with the indicated concentrations of CPT (0, 3, 6, and 12 μM) for 24 h. *P < 0.05, **P < 0.01, and ***P < 0.001 versus the control group. (B) Western blot analysis of LC3-I conversion to LC3-II in Huh7 and MHCC97-H cells treated with 6 μM CPT in the absence or presence of 10 μM chloroquine (CQ) or 5 mM 3-methylamphetamine (3-MA) for 24 h was conducted. *P < 0.05, **P < 0.01, and ***P < 0.001 versus the CPT group. (C) Huh7 and MHCC97-H cells were treated with or without 6 μM CPT for 24 h. LC3B puncta were then stained with anti-LC3B antibody by immunofluorescence and imaged with a confocal microscope. (D) Cells were treated with or without 6 μM CPT for 24 h and the changes in ultrastructure were detected by transmission electron microscopy. Arrows: autophagosomes/autolysosomes. Scale bar: 1 µm.