Figure 5
Parental RNAi of Tctra affects splicing of both Tcdsx and Tctra pre-mRNA.
(A) Multiplex PCR with Y-specific and non sex-specific primers amplifies two bands in males whereas only one band in females. About 9% of adults developed from eggs laid by females injected with Tctra dsRNA showed male phenotype during the pupal stage (the presence of male papillae) and the adult stage (the presence of male-specific bristles) amplified a single band similar to that detected in control females. All the other adults (~91%) were genetic males since two bands (as in control males) were detected. (B) Genetic males developed from eggs laid by either malE or Tctra dsRNA injected females showed male-specific Tcdsx isoform (Tcdsxm). The rare genetic females (escapers), developed from Tctra dsRNA injected females, also showed Tcdsxm isoforms. (C) Genetic males developed from eggs laid by either malE or Tctra dsRNA injected females showed male-specific Tctra (Tctra-m). The genetic females (masculinized) developed from eggs laid by Tctra RNAi females showed both Tctram and Tctraf isoforms. (D) Amplification product of rp49 shown as a control. (E)–(H) The ovaries from Tctra (E&F) or malE (G&H) dsRNA injected adults were dissected on 20th day after injection of dsRNA, stained with acridine orange and photographed using a fluorescent microscope. F&H show higher magnification image of a single ovariole.
