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Fig. 4. Defective pole plasm assembly in vls. (A,B) Absence of pole cell formation in vls³. Staining of Vas with rat anti-Vas antibodies (red) and DNA with Oli-Green (green) in wild-type (A) and vls³ (B) blastoderm embryos. (C,D) vls suppresses ectopic anterior pole cell formation induced by osk-bcd 3'UTR. Staining of Vas protein (red) and DNA (green) in osk-bcd 3'UTR (C) and vls³; osk-bcd 3'UTR (D) blastoderm embryos. No pole cells form at the anterior pole of vls³; osk-bcd 3'UTR embryos. (E,F) Osk is localized at the posterior pole of the oocyte in vls³ stage 10 egg chambers, albeit in reduced amount. (E) Wild-type and (F) vls³ egg chambers. (G) The level of Osk protein is reduced in vls. Ovarian protein extracts of wild-type (WT), vls¹, aub^N11/aub^HN2 and osk⁸⁴/Df(3R)pXT103 females were separated by SDS-PAGE electrophoresis, blotted and detected using anti-Osk antibodies. The abundance of short Osk is reduced in vls mutant ovaries, whereas the long form is maintained at the wild-type level. Both Osk forms are strongly reduced in aub ovaries, with a more substantial effect on short Osk. In contrast to aub, the phosphorylation of short Osk is not affected in vls. The same blot was successively probed for ribosomal P40, as loading control (Török et al., 1999). (H,I) Vas is localized at the posterior pole of the oocyte in wild type (H) and vls³ (I) stage 10 egg chambers. There is less localized Vas in vls³ compared with wild type. Ovaries were stained with rabbit anti-Vas antibodies (red) and Oli-Green for DNA (green). (J,K) Tud is detected in nuage and at the oocyte posterior pole in wild-type stage 10 egg chambers (J) but is not detected at these locations in vls³ stage 10 egg chambers (K). Ovaries were stained with anti-Tud antibodies (red) and Oli-Green for DNA (green).

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