γ-Tubulin is not required to recruit SAS-6 or SAS-4 to centrioles but is required for stable incorporation of SAS-4 during late prophase. (A) Kinetic profiles for recruitment of GFP:SAS-6 in γ-tubulin–depleted embryos (tbg-1[RNAi]) generated as described in Fig. 2 (B and C). (B) Kinetic profiles for the recruitment of GFP:SAS-4 in embryos depleted of γ-tubulin (tbg-1[RNAi]) or simultaneously depleted of γ-tubulin and SAS-5 (tbg-1,sas-5[RNAi]), generated as described in Fig. 3 A. (C) Levels of centriolar GFP:SAS-4 in γ-tubulin–depleted embryos, calculated by subtracting the GFP:SAS-4 signal in tbg-1,sas-5(RNAi) embryos from the combined centriole and PCM signal in tbg-1(RNAi) embryos, compared with centriolar GFP:SAS-4 in control embryos (control data are from Fig. 3 D). (D) High-resolution images of control and tbg-1(RNAi) embryos, generated by mating as in Fig. 1 A, expressing GFP:SAS-4 or GFP:SAS-6. Times (in seconds relative to cytokinesis onset) correspond to meiosis (−1,305 to −1,650 s), early (−1,166 to −1,002s) and mid (−698 to −482 s) S phase, and late prophase (−320 to −115 s). (E) Analysis of the central and peripheral GFP:SAS-4 signal in prometaphase/metaphase tbg-1(RNAi) embryos performed as in Fig. 3 F. Asterisks denote statistically significant differences relative to control (P < 0.05 by t test). (F) Two representative examples of photobleached centrosomes in late prophase/prometaphase control (top and bottom centrioles were bleached at 215 and 332 s before cytokinesis onset, respectively) or tbg-1(RNAi) (top and bottom centrioles were bleached at 321 and 32 5s before the onset of cortical contractility, respectively) embryos. Times are in seconds after photobleaching. All error bars indicate the 90% confidence interval. Bars, 5 μm.