siRNA-transfected U-2-OS or MDA-MB-231 cells were seeded onto six-well plates in technical triplicates or on 6-cm plates in technical duplicates. After 8 to 14 days, cells were fixed and colonies were stained with crystal violet (Sigma-Aldrich). For colony formation capacity, graphs show the average of at least two biological replicates and SEM. For survival analysis upon drug treatments, MDA-MB-231 cells were seeded according to the plating efficiency. After 24 hours of plating, cells were treated with AZD7762 for 24 hours and then washed and grown in fresh medium or were treated with olaparib and left in the culture continuously. Cells were incubated at 37°C for 10 to 14 days. Colonies were fixed and stained as described above. The colonies were counted using an in-house–built ImageJ macro using a Trainable Weka Segmentation plugin. On the basis of the colony number, plating efficiency (PE = number of colonies formed/number of cells seeded) and surviving fraction (SF = number of colonies formed after siRNA treatment/number of cells seeded × PE) were calculated and plotted.

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