Fig. 4

Effect of derivative 12 on the inhibition of aurora kinases. HCT116 cells were treated either with various concentrations of derivative 12 (0, 50, or 100 μM) for 60 min (a) or 50 μM derivative 12 over a time-course (0, 60, or 120 min) (b). Western blot analysis was performed using phospho-specific antibodies against aurA (Thr288)/aurB (Thr232)/aurC (Thr198). The anti-GAPDH antibody was used as an internal control. The band intensities of phosphorylated (p)-aurA, (p)-aurB, and (p)-aurC relative to GAPDH level were measured using ImageJ software. The data are presented as means ± SD (n = 3). NS, not significant; ***, P < 0.001 according to Dunnettt's multiple comparisons test