Fig. 12

Illustrating example of the gating strategy used for neuronal nuclei sorting. Single nuclei were gated (R1) based on their DAPI labelling, followed by gating neuronal nuclei (NeuN+) and non-neuronal nuclei (NeuN-) on the NeuN vs red autofluorescence dot plot. Representative western blotting of isolated nuclei obtained by FACS of frontal cortex area 8 of middle-aged (MA) and Parkinson’s disease (PD) stage 5–6 cases. No signal was detected with anti-superoxide dismutase 1 (SOD-1) antibodies but there was one specific band at 17 kDa with anti-histone H3 antibody, thus demonstrating no cytoplasmic contamination of isolated nuclei. One band at about 20 kDa and two bands at about 50 kDa and at 100 kDa are detected with specific anti-α-synuclein oligomeric species in the neuronal nuclear sample (NeuN+) in PD. Interestingly, a weak α-synuclein low band is also detected in the non-neuronal nuclei sample in PD