Fig. 7. MIG-17 glycosylation and localization in cogc-3(k181) and cogc-1(k179) mutants. (A) Percentages of anterior and posterior gonad arms with DTC migration defects in cogc-3(k181) worms carrying extrachromosomal transgenic arrays. cogc-3 expression under the muscle-specific promoter unc-54 rescues the DTC phenotype of cogc-3 mutants. Both phase 2 and phase 3 defects were partially rescued (data not shown). The same transgenic array caused no DTC migration defects in wild-type background, indicating the partial rescue is not due to the overexpression of COGC-3. At least two independent lines were scored for transgenics, yielding similar results. Data for representative animals are shown. n=120 for each experiment. The error bars represent the mean±s.d. The values were calculated from the data collected from the first and second half of the animals examined. (B) N-glycosylation of MIG-17. Worm lysates were immunoprecipitated and immunoblotted with anti-GFP. The signals correspond to proforms of MIG-17-GFP. MIG-17-GFP in cogc-3(k181) and cogc-1(k179) mutants migrate further than wild-type MIG-17-GFP. Brackets indicate smearing of the bands in cogc-3(k181) and cogc-1(k179). The underglycosylation of MIG-17-GFP is more prominent (arrowhead) in cogc-1(k179). Part of each immunoprecipitate was treated with PNGase F (N-glycanase) and immunoblotted with anti-GFP. (C) Gonadal localization of MIG-17-GFP in cross sections (upper), and corresponding schematic diagrams (lower). Sections were stained with anti-GFP (red), fluorescein phalloidin (green) and DAPI (blue). The circular arrangement of nuclei in wild-type, cogc-3(k181) and cogc-1(k179) animals correspond to germ line nuclei of the gonads. Arrowheads indicate the gonadal localization of MIG-17-GFP to the surface of distal (upper) and proximal (lower) gonad arms in wild type. The gonadal localization is shown in red circles in the diagram. Dorsal is towards the top. Scale bar: 12.5 µm. (D) Quantitative analysis of MIG-17-GFP localization in wild type and mutants. n=60 for each experiment.