Fig. 2

Biochemical analysis of alpha-synuclein in patient brains. Biochemical analysis of alpha-synuclein in patient brains shows that SDS-extractable phosphorylated alpha-synuclein was only observed in samples from patient brains a but not in a control brain b. Sequentially extracted fractions from brain homogenates of the MSA1, MSA2, iLBD1, and iLBD2 cases contain similar monomeric and oligomeric species of alpha-synuclein when probed with the EP1536Y antibody against phosphorylated alpha-synuclein (upper panel) and with the Syn211 antibody against alpha-synuclein (lower panel). Both antibodies recognize a prominent band at 64 kDa that may represent a tetramer (arrow) a and that is absent in control brain b. Sequential extractions from a control brain did not reveal phosphorylated alpha-synuclein and only showed monomeric alpha-synuclein in brain homogenate (BH), high salt (HS), and high salt-triton (HS-T) fractions (arrow) but not in the RIPA or SDS fraction b. Molecular sizes are shown in kilodalton