Curcumin's Cell Cycle and Apoptosis Effects

Cell cycle and apoptosis analyses were performed as described previously [25 (link), 26 (link)]. EC9706 (1 × 10⁴/well) were seeded in 96-well plates and incubated overnight, and treated with different doses of curcumin (0, 4, 8, 12, 16 μM) for 24 h. For apoptosis detection, the cells were stained with Hoechst (Beyotime, Shanghai, China) and Annexin V–FITC Kit (Beyotime, Shanghai, China). For cell cycle analysis, the curcumin-treated cells were fixed with 4% paraformaldehyde and stained with DAPI (Solarbio, Beijing, China) according to the protocol and protected from the light for 20 min at 37 °C in a 5% CO2 incubator. They were analyzed by In CELL Analyzer 2000.