Figure S5.
The PP1 motif in PAR-2 is crucial for proper cell polarity. (A) Embryonic lethality of gfp::par-2° (refer also later to a 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) homozygous mutant strain. The reported values correspond to the percentage of unhatched embryos over the total progeny (larvae and unhatched embryo). gfp::par-2°, n = 2,469, gfp::par-2(RAFA), n = 1,028. N = 4. Mean is shown and error bars indicate SEM. The P values were determined using two-tailed unpaired Student’s t test. (B) Measurement of the PAR-2 cortical intensity in the wild-type gfp::par-2 and in the gfp::par-2° embryos at pronuclear meeting. Numbers inside the bars indicate sample size. Mean is shown and error bars indicate SD. The P values were determined using two-tailed unpaired Student’s t test. N = 2. (C) Quantification of the PAR-2 size domain in live zygotes at pronuclear meeting, comparing gfp::par-2 and gfp::par-2°. Numbers inside the bars indicate sample size. N = 2. Mean is shown and error bars indicate SD. The P values were determined using two-tailed unpaired Student’s t test. (D) Quantification of the cleavage furrow position in gfp::par-2°and gfp::par-2(RAFA). gfp::par-2(RAFA) mutant show cleavage furrow position more toward the anterior compared with gfp::par-2° (n = 23 and n = 17, respectively). N = 6. Mean is shown and error bars indicate SD. The P values were determined using two-tailed unpaired Student’s t test. (E) Quantification of the PAR-2 intensity from midsection images of live one-cell stage embryos of the indicated genotype. gfp::par-2, n = 28, gfp::par-2(RAFA), n = 22. Imaging of the gfp::par-2, used as control, has been performed the same days as the analysis of the gfp::par-2(L165V) shown in Fig. 5 B. N = 2. Mean is shown and error bars indicate SD. The P values were determined using two-tailed unpaired Student’s t test. (F) On the left, representative Western blot of embryo extract, showing level of GFP::PAR-2 and TUBULIN, which is used as a loading control, for both gfp::par-2 and gfp::par-2(RAFA). On the right, normalized ratio GFP::PAR-2/TUBULIN. N = 3. The P value (equal to 0.1871 [ns]) was determined using two-tailed paired Student’s t test between the values of gfp::par-2 and gfp::par-2(RAFA). n = number of embryos analyzed; N = number of independent experiments. Scale bars, 5 µm. Anterior is to the left and posterior to the right. ns, P > 0.05; ****, P < 0.0001. Source data are available for this figure: SourceData FS5.

The PP1 motif in PAR-2 is crucial for proper cell polarity. (A) Embryonic lethality of gfp::par-2° (refer also later to a 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) homozygous mutant strain. The reported values correspond to the percentage of unhatched embryos over the total progeny (larvae and unhatched embryo). gfp::par-2°, n = 2,469, gfp::par-2(RAFA), n = 1,028. N = 4. Mean is shown and error bars indicate SEM. The P values were determined using two-tailed unpaired Student’s t test. (B) Measurement of the PAR-2 cortical intensity in the wild-type gfp::par-2 and in the gfp::par-2° embryos at pronuclear meeting. Numbers inside the bars indicate sample size. Mean is shown and error bars indicate SD. The P values were determined using two-tailed unpaired Student’s t test. N = 2. (C) Quantification of the PAR-2 size domain in live zygotes at pronuclear meeting, comparing gfp::par-2 and gfp::par-2°. Numbers inside the bars indicate sample size. N = 2. Mean is shown and error bars indicate SD. The P values were determined using two-tailed unpaired Student’s t test. (D) Quantification of the cleavage furrow position in gfp::par-2°and gfp::par-2(RAFA). gfp::par-2(RAFA) mutant show cleavage furrow position more toward the anterior compared with gfp::par-2° (n = 23 and n = 17, respectively). N = 6. Mean is shown and error bars indicate SD. The P values were determined using two-tailed unpaired Student’s t test. (E) Quantification of the PAR-2 intensity from midsection images of live one-cell stage embryos of the indicated genotype. gfp::par-2, n = 28, gfp::par-2(RAFA), n = 22. Imaging of the gfp::par-2, used as control, has been performed the same days as the analysis of the gfp::par-2(L165V) shown in Fig. 5 B. N = 2. Mean is shown and error bars indicate SD. The P values were determined using two-tailed unpaired Student’s t test. (F) On the left, representative Western blot of embryo extract, showing level of GFP::PAR-2 and TUBULIN, which is used as a loading control, for both gfp::par-2 and gfp::par-2(RAFA). On the right, normalized ratio GFP::PAR-2/TUBULIN. N = 3. The P value (equal to 0.1871 [ns]) was determined using two-tailed paired Student’s t test between the values of gfp::par-2 and gfp::par-2(RAFA). n = number of embryos analyzed; N = number of independent experiments. Scale bars, 5 µm. Anterior is to the left and posterior to the right. ns, P > 0.05; ****, P < 0.0001. Source data are available for this figure: SourceData FS5.

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