Fig. 3.

Crenolanib impairs mitosis and prevents centrosome clustering. a Percentage of ECRF24 and HUVEC cells entering mitosis during the 24 hr live-cell imaging movies (mitotic index) and the percentage of cell death after treatment with 0.5, 2 and 5 µM of crenolanib or DMSO. b Plots for the timing between nuclear envelope breakdown and anaphase onset in minutes (mitotic timing). Number of cells are ECRF24: 0 µM n = 230, 0.5 µM n = 229, 2 µM n = 84; HUVEC: 0 µM n = 81, 0.5 µM n = 80, 2 µM n = 54. c Representative live-cell imaging stills over time of mitotic ECRF24 (top) and HUVEC cells stained with SiR-tubulin (microtubule marker) and treated with indicated doses of crenolanib. The arrows indicate the two daughter cells. d Quantification of multipolar spindles in mitosis in ECRF24 and HUVEC cells treated with indicated crenolanib concentrations. On the right, representative images of a mitotic ECRF24 cell (CTRL and crenolanib 2 µM) displaying multipolar spindles (indicated by arrows) are shown. The hashtag symbol indicates the centrosome clustering event and the asterisks the daughter cells resulting from the asymmetric division. Scale bars indicate 10 µm. Significance (*P < 0.05, **P < 0.01) is indicated as compared to control (0.1% DMSO) and error bars indicate SD