Fig. 4

Depletion of RCC2 impairs mitotic entry and prometaphase to metaphase progression. a RCC2 was knocked out in HeLa cells by CRISPR, and Western blot was performed with indicated antibodies (n = 3). b Wild-type and RCC2-depleted HeLa cells were incubated with Nocodazole (100 ng/ml) or Taxol (2 μM) for 12 h, harvested and analyzed using flow cytometry (n = 3). The percentage of mitotic cells positive for phospho-histone H3 (pH 3) is indicated. c Quantitative results representing the mean ± SD of three independent experiments. Error bars indicate SD. *, p < 0.05. PI, propidium iodide. d Time-lapse images showing prolonged prometaphase and metaphase progression in RCC2-depleted HeLa-H2B cells, compared with control. Scale bars, 2 μm; the unit for time is minute. e Quantification of mitotic cells showed in D (n = 15 mitotic cells per group) and results represent mean ± SD. Error bars indicate SD. *, p < 0.05. (F) A RCC2-deficient HeLa cell line stably expressing Flag-tagged RCC2 was generated. Quantification of mitotic cells showed in Additional file 1: Figure S1C (n = 15 mitotic cells per group) and results represent mean ± SD. Error bars indicate SD. *, p < 0.05. NS, not significant