Depletion of PCNT or augmin accelerates centriole separation in haploid cells. (A) IB of PCNT, HAUS6 in RNAi-treated haploid, diploid, and tetraploid HAP1 cells. β-Tubulin was detected as a loading control. (B) Flow cytometric analysis of FSC and DNA content in RNAi-treated haploid, diploid, and tetraploid HAP1 cells stained by Hoechst. Representative data from two independent experiments are shown in A and B. (C) Live imaging of RNAi-treated haploid GFP-centrin cells taken at 5-min intervals. Cytokinesis onset was set as 0 min. Broken lines mark cell boundaries. Centrioles (indicated by magenta boxes) are enlarged in the right panels. (D) Time course of intercentriolar distance in C. Values represent means ± SE of at least 14 cells from two independent experiments for each data point (at least 18 cells were analyzed for each condition). Green or magenta markers at the bottom of the graph indicate statistically significant differences between mock-depleted and PCNT-depleted, or HAUS6-depleted haploid cells, respectively (P < 0.05, t test). (E) Cumulative frequency of cells in which intercentriolar distance had reached 0.8 µm in C. At least 18 cells from two independent experiments were analyzed for each condition.