Fig. 7: SNCA-A53T hiPSC-derived neurons exhibit accelerated α-Syn seeding and mitochondrial dysfunction.

a, Characterization of cortical neurons using immunocytochemistry at day 70 of neural induction (C1: control; C2: control, iso-CTRL: isogenic control of SNCA-A53T mutant line 1; A53T 1: SNCA-A53T mutant line 1; A53T 2: SNCA_A53T mutant line 2). ai, Neuronal marker MAP2. aii, Cortical layer VI marker TBR1. aiii, Total α-Syn. Quantification is shown in Extended Data Fig. 10ai–aiii. bi,bii, Lysates from SNCA-A53T neurons contain oligomers that cause increased membrane permeability compared to control neurons (n = 3 independent experiments). ci–ciii, Application of AF-488 A53T and AF594-A53T α-Syn to cells results in higher intracellular FRET intensity (n = 4, 5 or 6 independent experiments). di,dii, There is lower Δψm in A53T-SNCA neurons than in control, measured by TMRM fluorescence (n = 5 or 6 independent experiments). ei,eii, Increased production of mROS measured by mitoTrackerCM-H2Xros in SNCA-A53T neurons (n = 7 or 9 independent experiments). fi–fiii, SNCA-A53T neurons exhibit a higher redox index than control neurons, indicating complex I inhibition (n = 8 or 11 independent experiments). Data from individual lines are present in Extended Data Fig. 10b. gi–giv, SNCA-A53T require lower concentrations of ferutinin for mPTP opening. Data from individual lines are present in Extended Data Fig. 10c (n = 6 or 8 independent experiments). hi,hii, SNCA-A53T neurons exhibit higher cell death than control neurons at day 80 (there was no difference in basal cell death at day 60 as shown in Extended Data Fig. 10d), which can be rescued by 0.1 mM mito-TEMPO (n = 5 or 9 independent experiments). Note: Data are represented as data ± s.e.m. (box). *P < 0.05, **P < 0.005 and ***P < 0.0005. Detailed statistical information is shown in Supplementary Table 1. See also Extended Data Fig. 10. a.u., arbitrary units; BF, bright-field.

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