Figure 4

CPE affects the phosphorylation state of DAT. A, COS-7 cells were transiently transfected with DAT or together with CPE by using electroporation, grown for 3–4 days, and was collected to be homogenized. The cells lysates was immunoprecipitated with anti-DAT antibody overnight. Subsequent western blotting of SDS-PAGE immunoprecipitaed samples with polyclonal anti-phosphoserine antibody and reimmunoblotting the membrane with anti-DAT antibody. B, Phosphorylation state of DAT affected the interaction between DAT and CPE. Striatal slices were pretreated for 30 min with 2 μM PMA before immunoprecipitation with the DAT antibody. Subsequent western blotting of SDS-PAGE immunoprecipitaed samples were incubated with anti DAT or anti CPE antibody.