Validation screen for PLK1 inhibitors. (A) PLK1 inhibitors used in primary and secondary screens along with Aurora B inhibitor barasertib are depicted with their respective structures and clinical advancement. (B) Layout of the validation screen assay. (C) Dot plot representation of volasertib, GSK461364, and barasertib IC₅₀ values (Log₂ nM) in normal hematopoietic cells (2 samples are CD34⁺ cord blood cells and 1 sample is CD34⁺ mobilized peripheral blood), NK, and CK AML samples. (D) Correlation analysis of inhibitory responses (Log₂ IC₅₀ in nM) across AML samples. (E) IC₅₀ values (Log₂ nM) of volasertib, GSK461364, and barasertib are compared in CK AML samples according to their TP53 status (WT vs altered). Medians are represented by a black line in dot plots. alt, altered; NK, normal karyotype; wt, wild type.