FIGURE 7.
Rescue of the mutant by different chemical and pharmacochaperones. HEK293 cells were transfected with cDNAs encoding wild-type SERT, SERT-PG601,602AA, SERT-RI607,608AA, and SERT-RII607–609AAA. Twenty-four hours after transfection 2% DMSO (A), 100 μm 5-HT (B), 10 μm imipramine (C), or 100 μm ibogaine (D). After an incubation for another 24 h, membranes were prepared; the binding was carried out with these membranes (100 μg/assay) in the presence of 1 nm [3H]imipramine as outlined in the legend to Fig. 5. Binding to membranes prepared from untreated control cells expressing wild-type SERT was set to 100% to normalize for interassay variation in transient transfections; this 100% value corresponds to 0.32 ± 0.11 pmol/mg. Data are means from three independent experiments carried out in duplicate; error bars indicate S.E. Note that all assays were done in parallel such that the control conditions are the same in each panel; the separation in individual panels is to render the data more accessible. Data are means from five independent experiments carried out in duplicate; error bars represent S.E. Differences were tested for statistical significance by paired t test (*, p < 0.05; ***, p < 0.001).