C443 is the major site of palmitoylation of the D2R.

<p>(A) Schematic representation of D2L showing the location of cysteine residue mutations. (B) FLAG-tagged WT-D2R or D2R mutants were transiently expressed in HEK-293T cells and expression in cell lysates was analyzed by Western blotting. Untransfected cells served as controls. (C) Normalized amounts of 15-HDYA-labeled D2Rs were IP’d and fluorescently imaged at 725 PMT (top) and 760 PMT (bottom). (D) Total D2R from IPs was analyzed by Western blotting. (E) Levels of palmitoylated D2Rs were normalized to the amount of IP’d receptor. Brackets indicate bands corresponding to D2R that were used for quantitation. The bar graph represents the average pixel density (as determined by ImageJ) from five separate experiments. Data were analyzed using a two-sided unpaired Student’s <i>t</i> test (expressed as ± SEM, <i>n</i> = 5, ***P < 0.001).