Fig. 5. Cytoplasmic polyadenylation complexes in Drosophila ovaries. (A-D) Immunoprecipitations (IPs) were performed in ovary extracts either in the presence of RNase inhibitor or in the presence of RNase A (RNase). Co-immunoprecipitated proteins were identified by western blot. Mock IPs were with preimmune serum for Wisp and PAP IPs, with rabbit serum for BicC IP and with an irrelevant monoclonal antibody for Orb IP. Extract (1/20) prior to IP was loaded. (E) In vitro interaction assays showing that Orb directly interacts with recombinant GST-Wisp(702-1373), but not with GST-Wisp(11-547) or GST alone. Orb was revealed by western blot. 1/10 of in vitro synthesized Orb before the interaction assay was loaded (input). Protein extract from 0.5 ovary was also loaded (upper panel). Recombinant and GST proteins are shown (input, bottom panel).