Tmrm mitochondrial membrane potential indicator

Mitochondrial superoxide detection was performed by incubating the hiPSC-derived-neurons in HBSS containing 2.5 μM MitoSox Red mitochondrial superoxide indicator (Thermo Fisher Scientific) for 15 min at 37°C and 5% CO₂. After incubation, cells were washed three times with HBSS. Mitochondrial membrane potential was measured using 50 nM TMRM mitochondrial membrane potential indicator (Thermo Fisher Scientific) for 30 min at 37°C and 5% CO₂. Mitochondrial morphology was determined by staining the cells with 1 μM MitoTracker Green FM (Thermo Fisher Scientific) for 15 min at 37°C and 5% CO₂. Images were acquired in live using a Leica SP8-X confocal microscope (Leica Microsystems) and analyzed using Imaris software (Oxford Instruments).

Mitochondrial superoxide detection was performed by incubating cells in HBSS containing 2.5 µM MitoSox Red mitochondrial superoxide indicator (Thermo Fisher Scientific) for 15 min at 37 °C and 5% CO₂. Mitochondrial membrane potential was measured using 80 nM TMRM mitochondrial membrane potential indicator (Thermo Fisher Scientific) for 30 min at 37 °C and 5% CO₂. After incubation, cells were washed three times with HBSS. iPSC-derived neurons were imaged at day 40 of differentiation. Images were acquired in live conditions using a Leica SP8-X confocal microscope (Leica Microsystems) and analyzed using Imaris software (Oxford Instruments).