2.1.3. Cell Apoptosis

Annexin V-fluoroisothiocyanate (FITC)/propidium iodide (PI) double staining (Dojindo, Japan) was used to determine cell apoptosis. Cells were seeded at 1 <sup>×</sup> 10<sup>6</sup> cells per well in 6-well plates and cultured in growth media for 24 h. Subsequently, the medium was replaced with fresh growth medium containing different concentrations of glucose and curcumin as described above. After 48 h, cells were stained with PI and Annexin V-FITC in a dark room for 15 min, according to manufacturer instructions, and analyzed by flow cytometry (Cytomics FC 500 MCL, Beckman Coulter, Brea, CA, USA).
