Figure 2

AA005 induces non-canonical apoptotic cell death. (A) SW620 cells were treated with or without 20 μM camptothecin, 1 μM AA005, or 500 μM MNNG, for 36 h, 48 h or 8 h. Images were viewed using an Olympus BX-51 fluorescence microscope. Treatment with MNNG and camptothecin were used as controls. Scale bars: 100 μm. (B) SW620 cells were treated with 1 μM AA005 for 24 h or 48 h. Annexin-V/PI double-stained cells were measured by flow cytometry. Panels show percentages of Annexin-V⁺ or PI⁺ cells. Right: dead cells. Each column represents mean ± S.D. from an independent experiment performed in triplicate. **P < 0.01, compared with vehicle-only control. (C) TUNEL analysis of SW620 cells treated with 20 μM camptothecin, 1 μM AA005 and 500 μM MNNG. Images were viewed using an Olympus BX-51 fluorescence microscope. Scale bars: 20 μm. (D) Percentages of TUNEL⁺ cells were determined in 200 cells per sample and assessed in 3 independent experiments. Results show mean ± S.D. *P < 0.05, **P < 0.01, compared with vehicle-only control. (E) SW620 cells were incubated with 20 μM camptothecin, 1 μM AA005 or 500 μM MNNG for 36 h, 48 h or 8 h. DNA samples from these treated cells were electrophoresed in a 2% agarose gel.