Figure 4

17-DMAG is synergistic with cisplatin for Brca1 parental and cancer stem cells. Cells were treated with increasing concentrations of cisplatin, 17-DMAG (17-dimethylaminoethylamino-17-demethoxygeldanamycin hydrochloride), or a combination of both drugs at a constant 1:3 ratio. (a) Simultaneous treatment for 48 hours. The open circles and dotted lines represent exposure to 17-DMAG, the filled circles and dotted lines represent exposure to cisplatin as single agents, and the filled squares linked by a solid line show combination. (b) Sequential addition of the drugs. The filled circles show exposure for 48 hours to cisplatin, the open circles and dotted lines represent 24 hours pf exposure to 17-DMAG, and the filled squares linked by a solid line show combination. The ordinate indicates concentrations of cisplatin, and the error bars represent ± standatd deviation from sextuplicates wells of one of more than three independent experiments. (c) Visual representation of Combination Index (CI) for each combination calculated using CalcuSyn Software. The closed circles represent simultaneous treatment and the open circles represent sequential addition of cisplatin followed by 17-DMAG. Values of CI below 1 indicate synergy, values above 1 represent antagonism, and CI = 1 corresponds to an additive effect. (d) 17-DMAG sensitizes parental and cancer stem cells to Cisplatin. CI for cisplatin and 17-DMAG added simultaneously (+) or sequentially (→) to parental cells or cells sorted for putative cancer stem cell markers. Data are derived from more than three independent experiments.