Fig. 3
AZD7648 and doxorubicin have synergistic combination activity in breast and ovarian cancer cell lines. a Western blot analysis of OAW42 cells treated with doxorubicin (100 nM) ± AZD7648 (3 μM). b Concentration-dependent response to AZD7648 ± doxorubicin was measured by a Live/Dead assay. Graphs represent percentage viable cells ± SD following 5 days’ treatment relative to DMSO vehicle-treated controls from a representative experiment (n = 3). c High-content imaging analysis of indicated DNA damage markers in OAW42 cells. Cells were treated with increasing AZD7648 concentrations ± doxorubicin (3 nM) and immunofluorescently stained at indicated time points. Graphs represent mean ± 2 SEM γH2AX signal intensity, number of 53BP1 foci, number of cells/well, or micronuclei per cell from three independent experiments. Representative images are shown for AZD7648 (1 μM) ± doxorubicin (3 nM) at 48 h (scale bars, 25 μm). D represents DMSO vehicle-treated controls. d Synergy scores for the AZD7648 and doxorubicin combination in a panel of four ovarian and seven breast cancer cell lines. Cells were treated for 5–7 days and viability was measured by the Live/Dead assay. A synergy score of >5 is indicative of synergistic activity. e Activity heatmaps from representative experiments for MDA-MB-468, OAW42 and MDA-MB-436 cells. Experimental activity heatmap represents growth inhibitory (0–100) and cytotoxic activity (100–200) following treatment. Loewe additivity model fit heatmap represents expected activity values for an additive combination. Concentrations where combination activity occurred in excess of the expected activity are boxed in pink. Synergy scores from the representative experiment are indicated in brackets
