Fig. 3. F1-7 induced colon cancer cells apoptosis and ferroptosis.

a Colony forming assay of colon cancer cell lines. Cells were incubated with different concentrations of F1-7 for 24 h. On day 7, colonies were fixed and photographed. b, c Colon cancer cells were treated with F1-7 and AZD4547 for 48 h. Cells were stained with Annexin V and propidium iodide (PI), and then analysed by flow cytometry. Samples were measured in triplicate and experiments were independently repeated three times d Cells were incubated with F1-7 at different concentrations as indicated for 24 h, the cell lysates were prepared for western blot analysis to determine protein expression of cl-PARP, Bcl-2, Bax and GAPDH. e PI staining of HCT-116 cells with and without f1-7 with inhibitors of apoptosis and ferroptosis. The positive of cells were calculated. Data are expressed as mean ± SD (n = 3).