Figure 6.
PD98059-induced internalization of FLAG-hDAT is clathrin-mediated. FLAG-hDAT cells were transiently transfected with the neuronal-specific GFP-clathrin light chain. Double-staining experiments (red fluorescence for the FLAG-hDAT, green fluorescence for clathrin) were performed to determine colocalization of FLAG-hDAT and GFP-clathrin. Main panel, Confocal microscopy revealed little or no puncta of colocalization under control conditions. Inset, In FLAG-hDAT cells exposed for 30 min to 50μm PD9805, there is extensive colocalization (yellow) between GFP-clathrin and FLAG-hDAT, presumably at the level of endocytotic compartments.