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Figure 2


Fig. 2. DEPS-1 associates with P granules. (A) The predicted Y65B4BL.2/deps-1 gene and locations of mutant alleles. Coding region is in gray and UTRs are in white. In blue is a 61 amino acid region containing 21 serines, 7 threonines, 8 arginines and 7 alanines. bn121 removes nucleotides 988-1172 (horizontal line) and results in a predicted transcript with a stop immediately after amino acid M246. bn113 mutates a conserved residue in the 3' splice site. Amino acid and nucleotide sequence positions are with respect to the predicted translation start site. (B) Western blot analysis using affinity-purified anti-DEPS-1 and anti-{alpha}-tubulin as a loading control. The arrowhead indicates the position of DEPS-1. The asterisk indicates a ~120 kDa band present in wild type and in deps-1 mutants. Molecular mass of protein standards in kDa is on the right. (C-F) Anti-DEPS-1 staining pattern of wild type (C,D,F) and deps-1(bn121) (E) embryos at advancing stages of development (C, four cell; D,E ~16 cell; F ~100 cell). (G,H) DEPS-1::GFP is concentrated on P granules in ~100-cell embryos (G) and in germ lines and oocytes (H). (I-N) DEPS-1 and PGL-3 co-localize on P granules in wild-type germ lines (I-K, surface of the pachytene region) and oocytes (L-N). Merged images show DEPS-1 in green and PGL-3 in red. Arrowheads and arrows indicate P granules and nuclei, respectively. H-K are single-section confocal images; all other images are confocal stacks. Scale bars: 10 µm. Images C-G and I-N use the scale bar in C.





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