Figure 2. Kinetic profiles for SAS-6 recruitment in control,
sas-5(RNAi),
sas-4(RNAi), and HU-treated embryos. (A) High-resolution images of GFP:SAS-6 recruited to RFP-labeled sperm centrioles in embryos generated using the mating scheme in Fig. 1 A. Times (in seconds relative to cytokinesis onset) correspond to meiosis (-1,470 to -1,370 s), early (-1,180 to -1,156 s) and mid (-1,005 to -972 s) S phase, and late prophase (-281 to -267 s). (B) Normalized individual measurements of the integrated GFP:SAS-6 intensity coincident with sperm centriolar RFP signal (see Materials and methods for details on normalization). (C) Kinetic profiles for the recruitment of GFP:SAS-6 in control and
sas-5(RNAi) embryos. Data points are the mean of the normalized GFP intensity measurements collected during the 200-s interval centered on that point. (D) Depletion of SAS-4 does not affect the kinetics of centriolar GFP:SAS-6 recruitment or loss. (E) Recruitment profile of centriolar GFP:SAS-6 in embryos treated with 75 mM HU. The profile was generated as in C and D except that sequences were time-aligned with respect to centriole separation, which occurs at the same time after meiosis II anaphase in control and HU-treated embryos (Videos 1 and 2, available at
http://www.jcb.org/cgi/content/full/jcb.200709102/DC1). The control dataset plotted here is the same as in B-D except that the means were recalculated for the indicated 200-s intervals after converting seconds before cytokinesis onset to seconds after centriole separation. All error bars indicate the 90% confidence interval.