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In undisturbed and DNA-damaged cells, depletion of NEK11 prevents proteasome-dependent CDC25A degradation. NEK11 directly phosphorylates CDC25A, and CHK1 (checkpoint kinase 1) is activated directly by phosphorylation of NEK11 at Ser273. NEK11 is an important component of the G2/M checkpoint pathway, and genetic mutations in NEK11 may contribute to cancer development.