Fig. 1: Development and characterization of d-MSNs.

A Representative immunofluorescence images of d-MSNs from control and patient PKAN_[Tyr190*] differentiated for 42 days and stained with the neuronal markers microtubule associated protein 2 (MAP2), dopamine- and cyclic AMP-regulated phosphoprotein 32 (DARPP32), gamma-aminobutyric acid (GABA), glutamic acid decarboxylase 65/67 (GAD65/67), tyrosine hydroxylase (TH), vesicular glutamate transporter 1 (V-Glut1) and the astrocyte marker glial fibrillary acidic protein (GFAP). Nuclei were stained with Hoechst. Scale bars 20 μm. Histograms on the right show the percentage of cells positive for the indicated markers. All data are presented as the mean + SD, n = 3. B Representative immunofluorescence images of d-MSNs from the control subject stained with MAP2 and DARPP32 with enlargements to highlight the presence of spines in neuronal processes. Scale bars 20 μm. C Representative current-clamp recordings showing action potentials elicited in control and PKAN d-MNSs by injection of suprathreshold current steps. The plot on the right represents the fraction of total cells displaying repetitive firing (p = 0.7, Z score for 2 population proportions). D Examples of spontaneous IPSCs recorded in control and PKAN d-MNSs bathed in extracellular medium containing 5 μM NBQX to block glutamatergic AMPA receptors. Right, summary plot with fractions of total cells displaying IPSCs (p = 0.9, Z score for two population proportions). E Representative immunofluorescence images of d-MSNs from control and patient PKAN_[Tyr190*] differentiated for 42 days in the presence or absence of 20 μm CoA stained with MAP2, DARPP32 and Hoechst. The left histogram shows the number of counted cells per field. Data are presented as the mean + SD, n = 3, one-way ANOVA, *p < 0.05, **p < 0.01. The right histogram shows the percentage of cells positive for the DARPP32 marker. Data presented as the mean + SD, n = 3.