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.