Fig. 3. Loss of Hedgehog produces ovaries with predominantly cytoplasmic Sxl. (A-F) ovaries from hhts/hhAC females kept at 29°C for 8-9 days. (A) Germarium showing mostly cytoplasmic Sxl. 98.9% of hhts/hhAC ovarioles contain multi chambered cysts (n>50). (B) Germarium from a hhts/hhAC ovary stained with anti-Ptc antibody. The level of Ptc is high and the normal decrease in expression is absent (82%). (C) Germarium stained for -Spectrin. Branching of the fusome (arrow) occurs closer to the anterior end of the germarium than in wild type (Fig. 1C). (D) Germarium stained for Orb. (E) Germarium in D stained for Orb (red) and DNA (green). Orb accumulation shown with arrowhead. (F) Later stage egg chamber that lacks Sxl. This chamber is adjacent to the germarium in A; note brighter staining cells on left of panel. (G) Egg chamber from hhts/hhAC females with a constitutive allele of Sxl (SxlM4) stained for Sxl. Sxl expression is maintained in the later stage germ cells and remains cytoplasmic throughout. 87% of ovarioles contained multi chambered cysts (n>50). The nuclei in the chambers (F,G) are polyploid. (H) RT-PCR of RNA from ovaries of hhts/hhAC females, females with a transgene of the hedgehog cDNA under a heat-shock promoter (hs-hhM11), Sxlf4 and wild-type females. Significant levels of male Sxl RNA are detected in Sxlf4, hhts2/hhAC and hs-hhM11 ovaries.