The centrosome separation defect of KLP-7–depleted embryos is suppressed in plk-1(or683ts). (A) Still images from time-lapse recordings of AB and P1 cells from embryos of the indicated genotype. ZU655 (containing the plk-1(or683ts)) and its parental strain SA250 were used in this experiment. (B) Quantification of the centrosome separation ratios of the indicated genotypes. (C) Localization of KLP-7 in images of fixed embryos during the second division of the SA250 strain with or without klp-7(RNAi) and the ZU655 strain containing the plk-1(or683ts) allele with or without klp-7(RNAi). KLP-7 localizes to the centrosomes, kinetochores, and cytoplasm. (D) Tracking of the centrosome-to-centrosome distance over time in klp-7(RNAi) embryos (n = 11 AB cells and 10 P1 cells) and in plk1(or683ts);klp-7(RNAi) embryos (n = 11 AB cells and 10 P1 cells). T = 0 was set at NEBD for AB and P1, respectively. The green line indicates the minimal distance, under which it was not possible anymore to resolve the two centrosomes. Error bars represent SEM. (E) Images of control, par-3(RNAi), and par-2(RNAi) fixed early two-cell stage embryos expressing an endogenously tagged PLK-1::GFP. PLK-1 is observed at centrosomes, at the midbody, and in the cytoplasm. (F) Quantification of the absolute intensity of PLK-1::GFP in the cytoplasm of AB and P1 cells of control, par-2(RNAi), or par-3(RNAi) embryos. Error bars indicate SEM. Scale bars, 5 µm. The number of cells/embryos analyzed is indicated at the bottom of the graph and the statistical test used is indicated in Table S1. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.001.