Fig. 7. Sgk1 interferes with ENaC–Nedd4-2 interaction. Oocytes were injected with cRNA encoding FLAG-tagged ENaC, Nedd4-2-CS and Sgk1, as indicated. After overnight incubation with [35S]methionine, membrane fractions were lysed and immunoprecipitations performed with anti-FLAG antibodies. (A) Immunoprecipitated material analyzed by SDS–PAGE autoradiography and (B) western blotting using an anti-Nedd4-2 antiserum. Lysates were analyzed by western blotting using (C) anti-c-myc (i.e. Sgk1) and (D) anti-Nedd4-2 antibodies.