Fig. 5: Overexpression of pix-1 results in disruption and CRISPR repair of pix-1(gk299374) results in normalization of muscle cell boundaries.

a, b Western blot analysis showing overexpression of PIX-1 in the strain [pix-1(gk299374); sfIs20] in which the muscle cell boundary defect observed in the nonsense mutant pix-1(gk299374) has been rescued by an integrated array of wild-type pix-1 expressed from a muscle-specific promoter (see Fig. 1c). a A representative western blot is shown reacted with antibodies to PIX-1, and to the loading control histone H3. b Graphical summary of four independent western blot reactions for wild type vs. pix-1(gk299374); sfIs20. Means and standard errors of the means are shown. The two strains show statistically different levels of PIX-1 using a two-sided students t-test with p < 0.0001 (indicated by ****). Images of the entire western blots are provided in the Source Data Files. c This same array when expressed in a wild-type background [wild-type; sfEx61] disrupts the muscle cell boundary. d pix-1(syb2137gk299374) is pix-1(gk299374) after CRISPR/Cas9 was used to repair the nonsense mutation. Confocal images of anti-PAT-6 are shown and reveal that the muscle cell boundary is normal. This is further evidence that the muscle cell boundary defect is due specifically to mutation in the pix-1 gene. Arrowheads point to muscle cell boundaries visualized by immunostaining with anti-PAT-6. Each image is a representative image obtained from at least 2 fixation and immunostaining experiments, and imaging at least three different animals. Scale bars, 10 μm.