FIGURE 3.
Co-immunoprecipitation of D2L DAR with the NKAα1 subunit from transfected HEK293T cells. Proteins were extracted, immunoprecipitated (IP) with anti-FLAG agarose, electrophoresed, and immunoblotted (IB) as described under “Experimental Procedures.” HEK293T cells were transfected with 15 μg each of either FLAG tag alone, FLAG-D2L receptor, or FLAG-D2L receptor with NKAα1 subunit. Left panel, the blot was probed with a polyclonal anti-D2S/L antibody and visualized using ECL after incubation with an HRP-conjugated anti-rabbit secondary antibody. Right panel, the blot in left panel was stripped of all antibodies, re-probed with a monoclonal anti-NKAα1 antibody, and visualized by ECL after incubation with an HRP-conjugated anti-mouse secondary antibody. The arrow indicates the position of the NKAα1 subunit migrating at ∼95 kDa.