Figure 6.

Flavokawain (FKB) induces autophagy by inhibiting mTOR and activating Beclin‐1 in thyroid cancer cells. (A) ARO, WRO, and TPC‐1 cells were incubated with the indicated concentrations of FKB for 24 h. Levels of protein expression were analyzed by Western blot using antibodies against the indicated autophagy‐related proteins and GAPDH. The number under each band represents the fold change of band intensity relative to control group. (B) WRO cells stably transfected with GFP‐LC3B were further transfected with Beclin‐1 siRNAs for 24 h, then treated with 6 μg/mL FKB for another 24 h. Cells were then examined by a fluorescence microscopy. Percentages of cells with more than four GFP‐LC3B dots were quantified. (C) WRO cells were transfected with Beclin‐1 siRNA or NC siRNA for 24 h, then treated with 6 μg/mL FKB for another 24 h. Levels of protein expression were analyzed by Western blot using antibodies against Beclin‐1, LC3B, and GAPDH. All data are expressed as the mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001