Fig. 1.

Insoluble αSyn was phosphorylated at Tyr136 as well as Ser129 in the DN-LBD brain. a Amino acid sequence of human αSyn. C-terminal phosphorylation sites are shown in bold and are underlined. b–f Brain lysates from DN-LBD (cases #1, #2, and #3), Li-LBD (cases #1 and #2), AD, and CJD patients were analyzed by SDS-PAGE followed by immunoblotting with b anti-αSyn antibody D119, c anti-pS129-αSyn antibody ab51253, d anti-pY125-αSyn antibody, e anti-pY133-αSyn antibody, and f anti-pY136-αSyn antibody. Molecular mass markers are indicated in kDa on the left side of each panel. Arrows indicate the top of the gel. In d and e, WT r-αSyn (40 μg) was incubated in the presence of 100 ng/mL Syk (S52-10G; SignalChem Pharmaceuticals) and 200 μM ATP in 100 μL of reaction buffer (20 mM Tris–HCl, pH 7.5, 50 mM KCl, and 10 mM MgCl₂) at 37 °C for 1 h, and the sample containing WT r-αSyn (2 μg) was loaded as a positive control (PC)