“…After 24 h of subculture at 37 1C, U-937, HeLa, HRT-18, and SH-SY5Y cell lines as well as human peripheral MNC (10 6 cells/mL) were treated with extract (10-1000 mg/mL) for 6, 12 or 24 h. The treated cells were then washed with cold D-PBS, stained simultaneously with Annexin V-FITC (BD Bioscience, San Jose, CA, USA) and 7-AAD (BD Bioscience, San Jose, CA, USA), and analyzed by flow cytometry using an FACS Calibur instrument (Becton & Dickinson, San Diego, California, USA) and the strategies described elsewhere (de Lima et al, 2010) in which at least 10 5 events were acquired in the gated populations. Data were analysed using the WinMDI 2.8 software.…”