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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.
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