Haspin promotes Aurora B localization to KPCs and chromosome arms during ICRF-193 treatment in HeLa cells. (A) Representative immunofluorescent-stained images of pseudometaphase HeLa cells (nocodazole arrested) ± ICRF-193 ± CHR-6494 treatment for 45 min. CREST, red; Aurora B, green. (B) Classification of Aurora B staining pattern. (C) Quantification of average Aurora B signal intensity spanning chromosome arms. Each scan was normalized to the highest value. ICRF, n = 42 chromosomes from 35 cells; Haspin, n = 51 chromosomes from 27 cells. (D) Quantification of Aurora B signal intensity on chromatin. P values from Student’s t test. *, P = 0.04; **, P = 0.01; ***, P = 0.001; ****, P = 0.0004. a.u., arbitrary unit. (E) Live single-cell analysis of mitotic progression. Quantitation of time to anaphase and decondensation during drug treatments. Each vertical bar represents one cell, with each cell being scored for time to anaphase and decondensation from the start of the time course. (F) Representative immunofluorescent-stained images of pseudometaphase HeLa cells (nocodazole arrested) ± ICRF-193 for 45 min. CREST, red; SUMO2/3, green. The individual channels are provided for reference in Fig. S5 A. The plot shows classification of SUMO2/3 staining pattern. Bars, 10 µm. Error bars, standard deviation. The images are representative examples of those collected from three independent experiments.