Depletion of UHRF1 causes chromosome misalignment. (A and B) DU145 and PC3 cells with or without UHRF1 depletion were established by stable transfection with shRNA vectors for UHRF1 or control, and UHRF1 expression was assessed by Western blotting (A). The cell cycle distribution was analyzed by flow cytometry (B). (C–F) The nuclear DNA in DU145 and PC3 cells with or without UHRF1 depletion was stained with DAPI (scale bar, 20 μm; inset scale bar, 10 μm, C). The percent of mitotic cells was assessed according to nuclei morphology. n > 100 cells (D). The chromosomes that failed to congress at the metaphase plate are highlighted by red arrows (scale bar, 5 μm, E). The percentage of cells with unaligned chromosomes was assessed. n > 80 cells (F). (G and H) DU145 cells with UHRF1 depletion were transiently transfected with plasmids expressing UHRF1^ΔRING or UHRF1^WT, and the nuclear DNA was stained with DAPI (scale bar, 5 μm, G). The percentage of cells with unaligned chromosomes was assessed. n > 80 cells (H). The data for quantification in D, F, and H are from n = 3 independent experiments. Results are represented as mean ± SD (one-way ANOVA test); error bars represent SD. n.s., not significant; **, P < 0.01; ***, P < 0.001. Source data are available for this figure: SourceData F1.