Research Paper Volume 13, Issue 23 pp 25365—25376

Figure 5. Src promotes synergistic effects of AB4 with 5-FU treatment. (A) HCT116/FU cells were transfected with His-Src or vector plasmids, followed by 25 μM AB4 and 20 μM 5-FU treatment. Bosutinib was used for Src signaling inhibition as indicated. Phosphorylated Src and cleaved caspase-9 protein were measured with Western blot assays. GAPDH was used as loading control. (B) Infected HCT116/FU cells were treated as indicated. Caspase-9 activity was evaluated in the indicated time. (C) MTT assays were performed with the infected HCT116/FU cells with gradient of 5-FU. HCT116/FU-vector cells were also treated with Bosutinib for Src signaling inhibition. (D) Infected HCT116/FU cells were treated with 25 μM AB4, 20 μM 5-FU and Bosutinib for 5 days. Cell viability was examined by MTT assays. (E) Infected HCT116/FU cells were treated with AB4,5-FU and Bosutinib. Cell apoptosis was analyzed by flow cytometry. (F) The percentage of LGR5⁺ of HCT116/FU-Src and control cells were analyzed by flowcytometry, which were also treated with 25 μM AB4 for 48 h. The results represent at least three independent experiments. Data represent as the means ± SEM. ^*P < 0.01.