Fig. 2

Mutations in the pita gene. a A schematic showing the constructs used to express wild-type and mutant variants of Pita in transgenic Drosophila lines. b Co-immunoprecipitation of CP190 with wild-type and CP190-interacting region-deleted Pita protein fused with 3×FLAG in S2 cells. Protein extracts from Drosophila S2 cells co-transfected with 3×FLAG-Pita and CP190 plasmids were immunoprecipitated with antibodies against CP190 (using nonspecific IgG as a negative control), and the immunoprecipitates (IP) were analyzed by western blotting for the presence of FLAG-tagged Pita proteins. The quality of immunoprecipitation was controlled by western blotting for the presence of CP190 protein. “Input” refers to samples of the initial protein extract; “output” refers to the supernatant after the removal of the immunoprecipitate (IP). c Western blot analysis (10% SDS-PAGE) of protein extracts from transgenic flies expressing wild-type and mutated variants of Pita