Fig. 4.

The effect of FX on autophagy induction in HepG2 cells. (a) FX induces time-dependent activation of the autophagy related proteins, LC3II and Becline-1. Western blot analyses were performed on the lysates of cells that had been treated with 30 μM FX for the indicated time period. β-actin served as a loading control. Protein levels were presented as relative band intensities to control (vehicle treated) group. Results represent the mean ± S.D. for four separate experiments. ** p < 0.01; *** p < 0.001. (b) The pictures of the fluorescence micrographs show the formation of AVOs resulting from the treatment of FX in HepG2 cells. Cells were incubated either in the presence or absence of 30 μM FX for 6 h and were stained with AO. The presence of AVOs was indicated by the red fluorescence. (c) Inhibition of FX-induced autophagy by C.C. Western blot analysis of Beclin-1 were performed with lysates of HepG2 cells that had been pretreated with 10 μM C.C for 1 h being followed by exposure to 30 μM FX for 1 h