Fig 1: Guidance receptors affect subcellular distribution and levels of the GTPase CED-10/Rac1.All embryos are oriented with anterior to the left and dorsal up. Boxed regions are amplified and enhanced equally for contrast. Number of embryos that showed the represented phenotype are indicated. (A) Embryos carrying an integrated rescuing ced-10::gfp (pjIs4) transgene were double-stained with antibodies to GFP (Abcam, ab6556) and to endogenous AJM-1 to indicate the adherens junctions [51], [52]. Dotted lines outline the basolateral region of the epidermal cells if the region is discernable. (B) Embryos carrying an integrated rescuing ced-10::gfp transgene were double stained with antibodies to GFP [49] and to endogenous basolaterally localized UNC-70/beta spectrin [33]. Readings were taken across two cell junctions using the line tool in ImageJ for both the UNC-70 and the GFP signal. 5 readings were taken per cell and averaged, and plotted in IPad Prism. SEM is shown. (C) Total levels of CED-10::GFP measured with antibody to GFP (Abcam, ab6556). Numbers below each lane are the levels of GFP normalized to HSP90 (Abcam ab13492) as loading control, and relative to wild type, averaged from 4 blots from two sets of lysates. (D) Genetic interactions of WAVE/SCAR and guidance receptor mutants with the integrated rescuing CED-10::GFP transgene. “Early Arrest” refers to embryonic arrest before morphogenesis begins. Asterisks indicate significant change in the phenotypes compared to the single mutants. * = p<.05, *** = p<0.001 as determined by a One-way Anova test followed by the Tukey test.
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