Figure 1
Functional characterization of dopamine (DA) response in U2OS-mock and U2OS-DAT cells in a TRACT assay. (a) FLAG-tag ELISA shows detection of C-terminal FLAG-tag of DAT, represented as mean ± SD of two separate experiments each performed in quintuplicate. (b) Representative xCELLigence growth curves after cell seeding, antagonist pretreatment and dopamine addition. (c) Representative vehicle-corrected xCELLigence traces of U2OS-mock and (d) U2OS-DAT cells after stimulation with increasing concentrations of dopamine. Data is normalized prior to agonist addition at time = 0 min. (e) Concentration-effect curves of dopamine on U2OS-mock and U2OS-DAT cells are shown as the net AUC of the first 120 min after stimulation normalized to the cell response of 31.6 µM dopamine. (f) Cell response of 10 µM dopamine (red bar, set at 100%) on U2OS-DAT cells pretreated for 1 h with 1 µM of one of following GPCR antagonists: SCH23390 (dopamine D1-like), raclopride (dopamine D2-like), doxazosin (alpha-1 adrenergic), yohimbine (alpha-2 adrenergic), propranolol (beta adrenergic). Data are shown as mean ± SEM of three to six individual experiments each performed in duplicate. Comparison of multiple mean values to vehicle control was done using a one-way ANOVA with Dunnett’s post-hoc test. *** p < 0.001.
