All experiments were carried out using a minimum of two biological replicates. No statistical method was used to predetermine sample size. No samples were excluded from the analyses. The experiments were not randomized, and investigators were not blinded to allocation during experiments and outcome assessment. The primary research objectives were to test the hypothesis that TLK activity was required for DNA replication. The resulting data led to the secondary hypotheses that TLK depletion would synergize with agents that exacerbate replication stress (namely, checkpoint and PARP inhibitors) and that TLK activity would be beneficial to many types of cancer cells. Experimental design was primarily controlled laboratory experiments using standard cell culture techniques. It also involved the retrospective statistical analysis of TCGA cancer data sets.

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