Fig. 4. Overexpression of Hh leads to fewer cells with cytoplasmic Sxl. All hs-hhM11 females heat shocked for 3 days. Germaria positioned with anterior (stem cells) to the left and are laser confocal images. (A) Germarium stained with anti-Sxl antibody. Only one or two cells show significant cytoplasmic Sxl (inset). Note that the germ cells show an overall decrease in Sxl (arrow) and signal is detected mostly from the surrounding follicle cells. 85% of germaria show a decrease of early germ cells with cytoplasmic Sxl. (B) Germarium stained with anti-Fu antibody. Fewer cells at the anterior end show significant Fu (85.5% germaria show decreased cells with Fu). Note that late germ cells show a significant decrease in Fu (arrow). (C) Germarium stained for Orb (green) and DNA (red). Orb expression begins in early to mid region 1 (arrow), as opposed to wild type, which begins in region 2 (see Fig. 1D). (D) Germarium stained for Ptc (red). (E) Ptc and nucleic acid (green). Ptc expression in the early germ cells (bar in D) is very reduced relative to wild type (82% of germaria show reduced levels of Ptc in the early germ cells). Somatic cells and later chambers are normal. (F) Germarium stained for -Spectrin. No spectrosome is visible in the early germ cells (arrow) and the fusome is also not detectable (87% of germaria have no visible fusome). Where reported, the percent for phenotype frequency is from a count of greater than 50 ovarioles.