FIGURE 10.

Co-immunoprecipitation of GRK2 and its mutants with D₂ DAR. A, HEK293T cells were transfected with FLAG-tagged D₂ DAR plus WT GRK2 or various GRK2 mutants as indicated. Solubilized cell lysates were immunoprecipitated (IP) using anti-FLAG-agarose, separated by SDS-PAGE, and immunoblotted (IB) with an anti-GRK2 antibody as described under “Experimental Procedures.” A representative experiment, which was performed twice with identical results, is shown. B, HEK293T cells were transfected with either FLAG-tagged WT D₂ DAR or FLAG-tagged GRK(−) D₂ DAR plus GRK2. Cells were treated with 200 ng/ml pertussis toxin (PTX) overnight or incubated in the absence (C) or presence of 10 μm DA for 30 min before harvesting. Solubilized cell lysates were prepared and subjected to immunoprecipitation using anti-FLAG-agarose as described in A. A representative experiment, which was performed three times with identical results, is shown. mut, mutant.