FIGURE 5.

AMPK does not form a macromolecular complex with the Na⁺/H⁺ exchanger NHE1 isoform. To detect whether AMPK can associate with NHE1, cell lysates were prepared from AP-1 cells stably expressing NHE1_HA and then incubated with rabbit IgG or polyclonal antibodies against the endogenous AMPKα1 and -α2 subunits. The immunoprecipitates (IP) were analyzed by SDS-PAGE and immunoblotting (IB) with mouse monoclonal anti-HA antibody to detect the core (cg) and fully (fg) glycosylated forms of NHE1_HA. Data are representative of two independent experiments.