FIGURE 4.

KLHL12 does not interact with the mouse D4 receptor. HEK293T cells were transiently transfected with plasmids expressing human HA D4.2 receptor (5.5 μg), mouse D4 receptor (not tagged; 5.5 μg), or Etag KLHL12 (5.5 μg), as indicated. 48 h post-transfection, cells were lysed, and 5% of the lysate was subjected to SDS-PAGE followed by immunoblotting (IB) with anti-D4DR (Chemicon) and anti-Etag to detect dopamine receptors or Etag KLHL12, respectively. The rest of the lysates were subjected to immunoprecipitation (IP) with anti-D4DR (Chemicon). Receptor-KLHL12 associations were examined through subsequent immunoblotting with anti-Etag. Immunoprecipitation of the receptors was confirmed by immunoblotting with anti-D4DR.