Fig. 3

TAAR1 signaling through PKA and RhoA occur in different intracellular sites. SK-N-SH neuroblastoma cell were transfected with DAT and either AKAR4 (a) or the Rho-FRET (b) sensors to which tags were added to the carboxy or amino termini to dictate localization to various subcellular domains (see Materials and methods). Cellular nuclei were stained with DAPI (blue). PKA activation by AMPH was detected in all compartments (c) but favored non-raft membranes. RhoA activation (d, f) was most robust near the ER (red). (*p < 0.05 and **p < 0.01 by one-way ANOVA with Dunnett’s multiple comparisons test compared with the highest responding compartment; n≥10 cells.)