Figure 4.

YFP–SERT–⁶⁰⁷RI⁶⁰⁸–AA fails to reach axonal compartments. Rat dorsal raphe neurons were transfected with plasmids encoding YFP–SERT (A) or YFP–SERT–⁶⁰⁷RI⁶⁰⁸–AA (B) using Lipofectamine2000. After 48 h, the neurons were fixed and stained for MAP-2. MAP-2 staining was detected using an Alexa Fluor 568-conjugated secondary antibody. Images were captured by confocal microscopy. YFP-tagged SERT was found in the MAP-2-negative compartment in 24 of 24 examined neurons; YFP-tagged SERT–⁶⁰⁷RI⁶⁰⁸–AA was confined to the MAP-2-positive compartment in 20 of 21 examined neurons. Data are from three independent experiments (i.e., 4 individual preparations of rat dorsal raphe neurons that were independently transfected in parallel with a plasmid encoding either YFP–SERT or YFP–SERT–⁶⁰⁷RI⁶⁰⁸–AA). Scale bars, 20 μm.