Fig. 2

Fluorescent photomicrograph of AO/PI double-stained WEHI-3cells. The cells were treated with BM at 14 μg/mL for 24, 48 and 72 h. a Control (untreated) cells showed a normal structure with no remarkable features of apoptosis and necrosis. b At 24 h, AO was interconnected with fragmented DNA (bright green) and the blebbing cells as an indicator of early apoptosis start to be seen. c An orange colour, representing late apoptosis, was noticed at 48 h. d At 72 h, the cells were orange with a blebbing membrane that indicated late apoptosis. At 72 h, secondary necrotic cells (bright red colour) were noticed. e The percentages of viable early and late apoptosis and secondary necrotic cells were increased after WEHI-3 cells were treated with BM in a time-dependent manner. Apoptosis increased significantly (*p < 0.05) in a time-dependent manner. VI: viable cells, EA: early apoptosis, LA: late apoptosis, SN: secondary necrosis