Effects of ecdysone signaling components on TF formation.

At ML3, TFs were stained with anti-En, confocal Z-stacks were acquired, and the number of TF stacks in ovaries of each genotype counted. An average of about two short TF stacks could be observed in wild-type ovaries. Precocious activation of the ecdysone signaling cascade either by removal of EcR or usp by RNAi from the somatic cells of the ovary, or by mis-expression of Br-Z2 and Br-Z3, results in more TF stacks at ML3. Very little change in TF stacks is caused by mis-expression of Br-Z4. ND: In Br-Z1 ovaries we cannot quantify TF stacks, since the anterior of the ovary is mis-organized and individual TF stacks are hard to distinguish. However, more TF cells are observed in Br-Z1 ovaries (Figure 5B). At LL3, TF stacks were observed using 1B1 labeling. On average, 17 or 18 TFs are formed in wild type ovaries. When large mutant clones of Eip75B07401 or ftz-f103649 are generated using C587-Gal4, UAS-Flp, less TF stacks form.