Subcellular purity and mitochondrial localization of α-synuclein
in human PD and normal brains. Western immunoblot analysis of marker
proteins for different subcellular fractions from substantia nigra (100 μg
of protein each) was carried out using antibodies to p97, TOM20, actin
(A), and Sec 61 and TGN 38 (B). C, amounts of
α-synuclein present in the mitochondrial fractions from substantia nigra
of normal and PD subjects as quantitated by ELISA. Values are mean ±
S.D. from three separate experiments. Western immunoblot analysis of
mitochondria (50 μg) using antibodies to α-synuclein of substantia
nigra (SN, D), striatum (ST, G), and cerebellum (CE,
J) and. F, I, and L are Western immunoblot analysis of
TOM20 levels (as a loading control) in D, G, and J blots,
respectively. E, H, and K represent the quantification of
α-synuclein immunoreactivity in Western blots D, G, and
J, respectively. Values are mean ± S.D. from three separate
experiments. *, p < 0.05 compared with corresponding
normal levels (one-way analysis of variance with Tukey-Kramer post test).
NS = normal subjects, N = nucleus, M =
mitochondria, MC = microsomes, and C = cytosol.