Figure 5.
Effect of APD treatment on the cellular expression of trafficking-impaired D2R mutant constructs. A, effect of APD treatment on the cell-surface expression of the trafficking-deficient D2R mutant, D2RW160A. HEK293T cells transiently transfected with cDNA for the FLAG-tagged D2RW160A mutant construct, FLAG-D2RW160A, were treated with the indicated APDs (10 μm, 24 h). The relative levels of cell-surface receptor were then quantified by probing intact cells with anti-FLAG antibody and are reported as a percentage of the signal from vehicle-treated cells. The signal for cell-surface receptor after treatment with all APDs except clozapine was significantly increased (n = 8, Tukey; p < 0.01 for aripiprazole, p < 0.001 for olanzapine, p < 0.0001 for all other APDs). B, effect of APD treatment on the total cellular expression of D2R mutant, D2RW160A. HEK293T cells transiently expressing a FLAG-tagged D2RW160A mutant construct were treated with the indicated APDs (10 μm, 24 h), and total cellular D2RW160A levels are reported as a percentage of the signal from vehicle-treated cells (n = 3). C, effect of APD treatment on the surface expression of the D2R glycosylation-deficient mutant, D2RN5,N17,N23→Q. HEK293T cells transiently expressing a FLAG-tagged D2RN5,N17,N23→Q mutant construct, FLAG-D2RN5,N17,N23→Q, were treated with the indicated APDs (10 μm, 24 h), and the relative levels of cell-surface receptor were quantified as in A. The signal for cell-surface receptor after haloperidol treatment was significantly higher than in untransfected cells or cells treated with either vehicle or clozapine (n = 16, Tukey, p < 0.0001). D, effect of APD treatment on the total cellular expression of D2RN5,N17,N23→Q. HEK293T cells transiently expressing the FLAG-tagged D2RN5,N17,N23→Q mutant construct, FLAG-D2RN5,N17,N23→Q, were treated with the indicated APDs (10 μm, 24 h), and total cellular D2RN5,N17,N23→Q levels are reported as a percentage of the signal from vehicle-treated cells (n = 16). Error bars, S.E.