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Functional characterization of the DAT and NET variants heterologously expressed in COS-7 cells.

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A, B. COS-7 cells were transfected with full length (FL) hDAT, hDATΔEX6, and empty vector pcDNA3 (Mock), and then subjected to a [3H]dopamine (DA) uptake assay (A) and [3H]WIN35,428 binding assay (B) in parallel. Two days after transfection, cells were incubated with 10 nM [3H]DA at 37°C for 6 min (A) or with 2 nM [3H]WIN35,428 at 4°C for 2 h (B) in the absence (Total) or presence (Nonspecific) of 100 µM cocaine (A) or 1 mM DA (B), respectively. C. COS-7 cells were transfected with FL hNET and hNETΔEX6, and then subjected to a [3H]NE uptake assay. Nonspecific uptake was determined in the presence of 10 µM nisoxetine. Values represent the mean ± SEM for 3 experiments each performed in triplicate. Statistical analyses were performed using an analysis of variance (ANOVA) with pair-wise comparisons by the Bonferroni method. *P<0.05 vs Nonspecific and/or Mock, #P<0.05 vs FL-DAT or FL-NET.

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2010-08-05
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