UHRF1 ubiquitination is critical for spindle architecture. (A) DU145 cells with EG5 depletion were transiently transfected with plasmids expressing EG5^WT or EG5^K1034R, and the nuclear DNA was stained with DAPI. Scale bar, 20 μm; inset scale bar, 10 μm. (B) The mitotic cells were identified according to nuclei morphology and the percentage of mitotic cells was assessed. n > 100 cells. (C) Chromosomes that failed to congress at the metaphase plate are highlighted by red arrows. Scale bar, 5 μm. (D) The percentage of DU145 cells with unaligned chromosomes was assessed. n > 50 cells. (E) Spindles were stained with immunofluorescent α-tubulin antibody and chromosomes were stained with DAPI. Scale bar, 5 μm. (F and G) Spindle pole distance was measured. n = 40 cells (F). The percent of cells at the metaphases with abnormal spindle geometry was assessed. n > 50 cells (G). The data for quantification in B, D, and G are from n = 3 independent experiments. Results are represented as mean ± SD; error bars represent SD. Dots represent individual cell samples in F; bars are median ± quartile. **, P < 0.01; ***, P < 0.001. One-way ANOVA test.