Fig. 3
Oligomers co-localise with ATP synthase. a Representative images of rat neuronal co-cultures treated with either monomers or oligomers over 7 days and probed with a filament specific antibody. The nucleus can be seen in blue (DAPI). b Representative immunocytochemistry images of rat neuronal co-cultures treated with oligomers. The cells were probed for ATP synthase subunit-α and filament α-synuclein. The nucleus can be seen in blue (DAPI). Scale bar = 10 μm. c Left: diffraction limited image of α-synuclein (scale bar = 5 µm). Right: super-resolved images of α-synuclein and ATP synthase and merged imaged (scale bars are 500 nm). Schematic diagram of DNA-PAINT: The secondary antibody conjugated with a docking strand binds the primary antibody. A fluorophore labelled complementary imaging DNA strand binds the docking strand on the secondary antibody allowing super-resolution imaging. d Representative PLA images of cells treated with either monomers, oligomers or no synuclein probing for ATP synthase subunit α and filament α-synuclein. Cultures were counterstained with the neuronal marker, MAP2. Scale bar = 10 μm. e Quantification of PLA signals per cell. N = 3 experiments. Oligomers: n = 182 cells; Monomers: n = 117; Control: n = 134 cells. f Representative PLA images of cells treated with oligomers probing for ATP synthase subunit α and filament α-synuclein. TOMM20 was probed to validate the mitochondrial localisation of the PLA signal. Two-tailed Student’s t-test for e. Scatter points represent number of puncta per cell. Data represented as mean ± SEM. ***p < 0.001
