Figure 3. Shikonin strongly induces late apoptosis in U87MG cells. A: U87MG cells were grown on 60 mm-diameter dishes. They were then treated with 5 nM vincristine, 2 μM shikonin, or 0.1% dimethyl sulfoxide (DMSO; Control). After 24 h, annexin V-propidium iodide (PI) analyses were performed. B: U87MG cells were grown on 60 mm-diameter dishes. They were then treated with 5 nM vincristine, 2 μM shikonin, or 0.1% DMSO (Control). After 24 h, staining with 4’,6-diamidino-2-phenylindole was performed. C: U87MG cells were grown on 60 mm-diameter dishes and treated with vincristine, shikonin or 0.1% DMSO (Control). After 24 h, fluorescence-activated sorting (FACS) analyses were performed. D: U87MG cells were grown on 60 mm-diameter dishes and treated with 5 nM vincristine, 15 μM compound 3K, or 0.1% DMSO (Control). After 24 h, FACS analyses were performed. E: U87MG cells were grown on 60 mm-diameter dishes. They were then treated with 15 μM compound 3K, or 0.1% DMSO (Control). After 24 h, annexin V-PI analyses were performed.