Depletion of PCNT or augmin accelerates centriole duplication in haploid cells. (A) Correlative live- and fixed-cell imaging. RNAi-treated HAP1 cells were live imaged by DIC microscopy every 5 min for 16 h (left) and then fixed and subjected to immunostaining against CP110 and SAS-6. The cells that passed the mitotic phase during time-lapse imaging were identified using grid patterns on the culture dish and observed using fluorescence microscopy (right). Anaphase onset was set as 0 min. (B) Immunostaining of CP110 and SAS-6 in RNAi-treated HAP1 cells in the correlative live and fixed cell imaging. Insets show 2× enlarged images of the centrioles in A and B. Broken lines mark cell boundaries. (C and D) Percentages of cells with indicated numbers of CP110 foci (C) or SAS-6 foci (D) at each time window after anaphase onset in B. At least 114 cells from two independent experiments were analyzed for each condition (at least 10 cells for each data point). Note that percentage of the cells that possessed three or four centrin foci increased in PCNT- or HAUS6-depleted haploid cells compared with control haploid cells in a 200- to 400-min time window, indicating that the initiation of centriole duplication was accelerated in these cells.