Effects of tozasertib on BCR-ABL-expressing cells. (A) The percentage of apoptotic cells was estimated by annexin V assay after culturing for 48 h with tozasertib. Results from 3 independent experiments were averaged. Error bars represent standard deviations. *P < 0.05, tozasertib treatment versus control in the same cell line. (B, C) K562 and Ba/F3 wt and T315I BCR-ABL cells were treated with tozasertib for 24 h. Total extracts were analyzed by immunoblotting with phospho-specific anti-Abl, -Crk-L, -cleaved caspase 3, and -cleaved PARP Abs. Actin was used as the loading control. Band intensities were quantified using ImageJ software. Data are representative of 3 separate experiments.