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Fig. 2. Oskar protein is ectopically expressed in the 18304 mutant. (A,E) oskar RNA localization in stage 10 wild-type and 18304 hemizygote mutant egg chambers revealed by in situ hybridization. oskar RNA localizes correctly at the posterior pole in wild-type (A) and 18304 mutant (E) oocytes. (B-D,F-H) Anti-Oskar immunostaining of wild-type and 18304 hemizygous egg chambers and embryos. (B,F) Stage 6 egg chambers of wild-type (B) and 18304 hemizygous (F) females. In the wild type, Oskar protein is not expressed before posterior localization of the RNA and is not detected at this stage (B), whereas it accumulates prematurely (arrow) in 18304 hemizygous oocytes (F). (C,G) Stage 10 egg chambers of wild type (C) and 18304 hemizygotes (G). In the wild type, Oskar protein is detected as a tight crescent at the oocyte posterior pole (C), whereas, in the 18304 mutant, it is observed all around the cortex of nurse cells and the oocyte (arrows), as well as at the posterior pole (G). The signal observed in the nuclei of wild-type and 18304 follicle cells, and in nurse cells is also observed in oskar protein-null egg-chambers (data not shown), indicating that it is background. (D,H) 0-2 hour-old embryo of wild type (D) and 18304 hemizygotes (H). In wild-type embryos, Oskar protein is maintained at the posterior pole as a tight crescent (D). In 18304 mutant embryos, Oskar protein is mislocalized all around the cortex, with a higher accumulation at the posterior pole (H). (I) Northern blot analysis of wild-type and 18304 hemizygote ovarian poly(A)+ RNA, probed with an oskar cDNA probe (top panel). No difference is detected in oskar RNA expression levels between wild type and the 18304 mutant. The blot was re-probed with rp49 cDNA as a loading control (bottom panel). (J) Western blot analysis of protein extracts of wild-type and 18304 mutant ovaries. Probing with anti-Oskar antiserum reveals that both isoforms of Oskar protein are expressed at higher levels in 18304 than in wild type (top and middle panels). The blot was re-probed with an anti-{alpha}-Tubulin antibody as a loading control (bottom panel).





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