Fig. 5
Impaired mitochondrial function in SNCA triplication iPS derived neurons. a Representative ICC images of control, isogenic control, and triplication cells probed for filament α-synuclein. The nucleus can be seen in blue (DAPI). b Representative PLA images and quantification of control and triplication iPS derived neurons showing a close proximity between filament α-synuclein and ATP synthase with the nucleus in blue (DAPI). TOMM20 was probed for to validate the mitochondrial localisation of the PLA signal. N = 3 experiments; control n = 155 cells, and triplication n = 115 cells. c Representative images of control and triplication iPS derived neurons loaded with TMRM and d ΔΨm quantification. N = 3 experiments, n ≥ 32 cells. e Representative traces of TMRM fluorescence in control and SNCA triplication neurons after addition of oligomycin (2 μg/ml), rotenone (1 μM), and FCCP (1 μM). f Representative trace of NADH in control and triplication iPS derived neurons. FCCP is applied to maximise respiration and therefore minimise the NADH pool and NaCN is added to block the mitochondrial respiration and therefore maximise the NADH pool. g Quantification of the redox index in control (N = 3 experiments, n = 112 cells) and triplication iPS derived neurons (N = 3 experiments, n = 148 cells). Two-tailed Student’s t-test for b, d, g. b Scatter points on scatter column represent number of puncta per cell. d, g Scatter points represent individual cells/areas. Data represented as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001
