Anthocyanins suppress tumor growth in a xenograft model and decrease cell growth and apoptosis in Hep3B hepato-carcinoma cells in vitro . (a) Hep3B hepato-carcinoma cells (1 × 106 cells/0.1 ml) were injected subcutaneously into the left flanks of Balb/C nu/nu mice (n = 5 per group). After 1 week, mice received anthocyanins s.c. (50 mg/kg/day) for 20 days. Tumor volume was measured once every 2 days and calculated as described in the Materials and Methods section. Body weight was measured once each week. *P < 0.05, compared with control tumor volume on day 18 or 20. (b) Cells were treated with anthocyanins (25–400 μg/ml) for 48 h, and cell viability was measured using the MTT assay. *P < 0.05 and ***P < 0.001, compared with control. (c) Anthocyanins-treated cells were stained with Annexin V-FITC, PI or Hoechst 33342 dye (10 μM) and analyzed by flow cytometry or fluorescence microscopy.