Mitochondrial targeting of WT/α-synuclein and
+33/α-synuclein in DAN neuronal cultures. DAN cells were transfected
with WT/synuclein-FLAG (A) and +33/synuclein-FLAG (D).
Cytosol (cyto.) and mitochondrial (mito.) fractions (100
μg) were probed with FLAG antibodies. B and C show the
levels of loading controls: TOM20 and actin, respectively, for A. E
and F show the levels of loading controls: TOM20 and actin,
respectively, for D. G, quantitation of WT/synuclein and
+33/synuclein in the mitochondrial and cytosolic fractions by ELISA at
different time points of transfection. Values are mean ± S.D. from
three separate experiments. *, p < 0.05 compared with
20-h transfection. H, following the transfection, the whole cell
lysates (homogenate) from DAN cells were tested for the levels of
α-synuclein at different time points using quantitative ELISA. Values
are mean ± S.D. from three separate experiments. No significant
variation was observed (one-way analysis of variance with Tukey-Kramer post
test).