Tetrandrine potently induces autophagy in a variety of cell lines. Western blot analysis of autophagy-related LC3 proteins in Hela, MCF-7 and HFF cells after treatment with various concentrations (A) and gradient times (B) as indicated. (C) Cells described in the Materials and Methods section were treated with rapamycin or tetrandrine alone for 24 hours. Autophagy-related LC3 proteins were analyzed by western blot to assess autophagy induction. (D) Treatment with 5 μM tetrandrine induces autophagy in Hela, MCF-7 and HFF cells as analyzed by the acridine orange staining assays described in the Materials and Methods section. AO fluorescence was observed with a fluorescence microscope. (E) Cells with GFP-LC3 dots accumulation exhibited autophagy after tetrandrine treatment and were viewed by confocal microscopy, Error bars represent ± SD, **p < 0.01.