Figure 2.

Astrocyte Co-culture Rescues Differentiation of hiPSC Line-Induced Human DA Neurons under KCN or Rotenone Treatment

(A) Representative images for immunostaining of MAP2 (green), TH (red), and Syn (white, color changed from far red) in induced DA neurons treated with KCN or rotenone with or without astrocyte co-culture. Scale bars, 25 μm.

(B) Quantification of neuronal process length of MAP2-positive neurons and TH-positive DA neurons using NIH ImageJ program.

(C) Quantification of numbers of Syn-positive clusters along the branches of hiPSC-derived DA neurons (TH-positive dendrites).

(D–G) Quantitative real-time PCR results of the indicated gene expression levels of AADC (D), DAT (E), VMAT2 (G), and MAP2 (F) in DA neurons with (+) or without (−) co-culture of astrocytes in the presence of vehicle, KCN, or rotenone.

Scale bars, 25 μm in (A). For (B)–(G), ^∗∗p < 0.01 versus the vehicle group. Statistical analysis was performed using StatView software (SAS Institute, v.5.0.1). One-way ANOVA was used for repeated-measure analysis, followed by Fisher's protected least significant difference for post hoc comparisons. Data are presented as means ± SEM. n = 3 independent experiments, with 12–13 cells quantified per experiment in (B) and (C). n = 3 independent experiments in (D)–(G).