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S1 Huh7 BEL7402 C Tet Rapa C Tet Rapa LC3-I LC3-II GAPDH Figure S1 Detecting Tetrandrine-induced autophagy in Huh7 and BEL7402 cell lines. Rapamycin treatment was used as a positive control. These cells were treated with 5μM Tetrandrine or 100nM rapamycin for 24 hours. Cell lysis were subjected to Western blot analysis to detect LC3 expression level.
S2 Huh7 BEL7402 HepG2 Figure S2 Acridine orange staining. (top) Huh7, BEL7402 and HepG2 cells were incubated with 5 μmol/L Tetrandrine for designated time points, and then stained with acridine orange (1 μg/mL)，and examined by a fluorescence microscope. Magnification, ×40. (bottom) The ratio of cells containing acidic vesicular organelles (AVOs). Columns, mean percentage of autophagy cells; bars, SD. At least 300 cells from each treatment group were observed.
S3 Huh7 Tet－＋－＋ 3-MA －－＋＋ LC3 II GAPDH Figure S3 Huh7 cells were pretreated with 2 mM 3-MA for 1 h, and then with or without 5 μM Tetrandrine for 24 hours, and the expression levels of LC3 were detected.
S4 Figure S4 Huh7 cells were treated with 5 μM Tetrandrine for indicated time. Cell lysis were collacted and subjected to Caspase 3 activity analysis. Level of untreated Huh7’s caspase 3 activity was set as 100%.
S5 Figure S5 Nude mice bearing Huh7 cell tumor xenografts were randomly distributed into two groups (n=7) and administered either the vehicle only (Vehicle) 25 or 50 mg/kg body weight tetrandrine by gavage for 20 days. The body weight of nude mice were measured every several days.
