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. 2008 Apr 25;283(17):11083–11096. doi: 10.1074/jbc.M708473200

FIGURE 1.

FIGURE 1.

KLHL12 interacts with the D4 receptor in eukaryotic cells. HEK293T cells were transiently transfected with plasmids expressing HA D4.2 receptor (5.5 μg) and/or Etag KLHL12 (5.5 μg), as indicated. 48 h post-transfection, cells were lysed. 5% of the lysates were subjected to SDS-PAGE and subsequent immunoblotting (IB) with anti-HA and anti-Etag to detect expression of HA D4.2 receptor and Etag KLHL12, respectively (bottom). The rest of the lysates were subjected to immunoprecipitation (IP) with anti-HA. The D4-KLHL12 association was examined through subsequent immunoblotting with anti-Etag. Immunoprecipitation of the HA D4.2 was confirmed by subsequent immunoblotting with anti-HA. *, this signal represents a combination of immunoglobulin G, a nonspecific binding signal from the anti-HA antibody, and a D4-specific signal in lanes 2 and 4. **, a combined nonspecific signal from anti-HA and a D4-specific signal in lanes 2 and 4.