Fig. 5

Effect of derivative 12 on the disruption of mitotic spindle dynamics. HCT116 cells were treated with 50 μM for 24 h. The cells were fixed and incubated with antibodies specific to mouse anti-α/β-tubulin (1:500) and rabbit anti-phospho (p)-Histone H3 (Ser10; 1:500) for 2 h, followed by incubation with AlexaFluor 488-conjugated (green signal) and AlexFluor 555-conjugated (red signal) secondary antibodies for 30 min. Nuclear DNA was stained with 1 μg/mL Hoechst 33,258 for 10 min (blue signal). Fluorescent cells were examined under an EVOSf1 fluorescence microscope. Int, interphase cell; Mit, mitotic cell; Size bar, 50 mm. Arrows, Mitotic spindles attached to metaphase chromosomes