Co-immunoprecipitation of NHERF1 with NPT2a in Hela cells.

<p>Hela cells were transfected with a plasmid containing the cDNA of NPT2a tagged with GFP at its N- extremity. Cells were also transfected with a plasmid containing the wild type cDNA of NHERF1 or one mutant (E68A or R153Q or E225K) tagged with Flag. Upper panel: Flag-tagged-NHERF1 protein was immunoprecipitated by using an anti-Flag antibody and the immunoprecipitates were probed with anti-GFP antibodies to detect GFP-tagged-NPT2a on Western blot. Middle panel: The total expression of GFP-tagged-NPT2a was analyzed in the cell lysates by anti-GFP Western Blot. Lower panel: Wild type and mutant Flag-NHERF1 was immunoprecipitated and probed by using anti-Flag antibodies to compare Flag-NHERF1 immunoprecipitation in the different conditions.