Figure 3.

Effect of Astrocyte Co-culture on Mitochondrial Functions in hiPSC Line-Induced Human DA Neurons under KCN or Rotenone Treatment

(A–C) Complex IV (A) and I (B) activity and ATP levels (C) were determined in hiPSC-derived DA neurons treated with KCN or rotenone with or without astrocyte co-culture. Data are expressed as fold change relative to the vehicle group in (A)–(C) (n = 3 independent experiments; mean ± SEM).

(D and E) Mitochondrial ROS levels were measured by MitoSOX staining. Representative images for MitoSOX staining for hiPSC-derived DA neurons with the above treatments are shown in (D), and quantification of MitoSOX staining intensity is shown in (E) (n = 3 independent experiments; mean ± SEM, with five cells quantified per experiment). Scale bars, 10 μm in (D).

(F and G) Assessment of intracellular ROS levels measured by EPR spectroscopy. Representative images for EPR are shown in (F), and quantifications are shown in (G) (n = 3 independent experiments; mean ± SEM).

^∗p < 0.05 versus the vehicle group (C, E, and G). Statistical analysis was performed using Statview software (SAS Institute, Version 5.0.1). One-way ANOVA was used for repeated measure analysis, followed by Fisher’s protected least significant difference for post hoc comparisons (A–C, E, and G).