Figure 1.

The effects of FAD on colorectal cancer cell lines. (a and b) Human colorectal cancer cells HCT116 and SW480 were treated with different concentrations of FAD for 48 h and cell viability was quantified. (c) Normal human colon epithelial cells FHC were treated with different concentrations of FAD for 48 h and cell viability was quantified. (d and e) HCT116 cells were treated with FAD for 48 h, and apoptosis was quantified by FACS after staining with Annexin V-FITC and propidium iodide. (f) Viability of HCT116 cells treated with 6 μM FAD for 48 h in the presence or absence of 100 μM Z-VAD-fmk. (g) Viability of HCT116 cells treated with 6 μM FAD for 48 h in the presence or absence of 40 μM Necrostatin-1 (Nec-1). (h) Viability of HCT116 cells treated with 3 μM FAD for 48 h in the presence or absence 20 μM of chloroquine (CQ). Trypan blue staining was used to determine cell viability. (i) HCT116 cells were treated with 6 μM FAD for the indicated periods, and LC3-I/II protein levels were determined by western blot analysis. (j) HCT116 cells expressing EGFP-LC3 were treated with 0 or 3 μM FAD for 30 h and were imaged with a fluorescence microscope. Serum-free (SF) treatment was used as a positive control for autophagy induction. *P<0.05 and **P<0.01