Fig. 1

Relation of Ser129-phosphorylated α-syn levels to total α-syn ones in intra- and extracellular spaces. CHO cells were transfected with the empty vector or the indicated amounts of wild-type α-syn cDNA. Samples were loaded along with recombinant α-syn proteins and Ser129-phosphorylated α-syn proteins for standards, followed by western blotting. Bands of Ser129-phosphorylated α-syn and total α-syn, including phosphorylated and non-phosphorylated forms, were detected by EP1536Y and Syn-1 antibody, respectively. Relative band intensities of Ser129-phosphorylated α-syn and total α-syn were corrected by plotting them on the standard curves, and then normalized to the intensities of β-actin. a Relation in intracellular α-syn. Cell lystaes (10 μg/lane) were loaded. b Relation in extracellular α-syn. After TCA-precipitated proteins were resolved by Laemmli’s sample buffer, samples corresponding to 20% of CM volume were loaded. Graphs show the positive correlation between Ser-129 phosphorylated and total α-syn levels