The PP1 binding site in PAR-2 is required in vivo for PAR-2 cortical posterior localization. (A) Representative midsection frames of time-lapse imaging of gfp::par-2° (the symbol ° indicates a mixture between wild-type gfp::par-2 homozygous worms expressing wild-type gfp::par-2 and heterozygous worms expressing wild-type gfp::par-2 from one allele and mutant gfp::par-2[RAFA] from the other allele) and gfp::par-2(RAFA) embryos at pronuclear meeting (n = 16 and n = 17, respectively). N = 6. (B) PAR-2 line profile of live zygotes at pronuclear meeting: gfp::par-2°, n = 16, and gfp::par-2(RAFA), n = 17; N = 6. Mean is shown and error bars indicate SD. (C) The wild-type and gfp::par-2(RAFA) dividing two-cell stage embryos. The upper panel shows a schematic representation, whereas in the lower panel images from Differential Interference Contrast (DIC) time-lapse movies are shown. In the schematics, the nucleus and the metaphase plate are in red, black lines are microtubules, and black circles are centrosomes. (D) Quantification of the cell cycle time in gfp::par-2° and gfp::par-2(RAFA) embryos. Numbers inside the bars indicate sample size. N = 6. Mean is shown and error bars indicate SD. The P values were determined using two-tailed unpaired Student’s t test. (E) Representative midsection frames of time-lapse imaging of gfp::par-2° and gfp::par-2(RAFA) one-cell embryos at pronuclear meeting. gfp::par-2°; ctrl(RNAi) n = 15, pkc-3(RNAi); gfp::par-2° n = 17, gfp::par-2(RAFA); ctrl(RNAi) n = 11, and pkc-3(RNAi); gfp::par-2(RAFA) n = 17; N = 7. (F) Quantification of PAR-2 intensity obtained as a ratio of PAR-2 posterior cortex intensity over the posterior cytoplasm. Numbers inside the bars indicate sample size. N = 7. Mean is shown and error bars indicate SD. The P values were determined using two-way ANOVA Tukey’s multiple comparisons test. ns, P > 0.05; **, P < 0.01; ****, P < 0.0001. In E and F, RNAi was performed by injection. n = number of embryos analyzed; N = number of independent experiments.