Development and characterization of hiPSC‐derived neurons from controls and PKAN patients Development and characterization of hiPSC‐derived neurons from.

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Development and characterization of hiPSC‐derived neurons from controls and PKAN patients Development and characterization of hiPSC‐derived neurons from controls and PKAN patients Representative IF image of NPCs stained for Nestin, FoxG1, and Pank2.NPCs differentiated into neurons by overexpressing Ngn2 (one representative experiment is shown). Two weeks after the infection differentiated NPC were positive for neuronal markers βIII tubulin (Tuj1), Map2, NeuN and human nuclei (hNu) and synaptic markers, the voltage‐gated Na+ channels (PanNav), and the vesicular glutamate transporter 1 (VGlut1).Western blot of soluble cell homogenates from human neurons probed with PANK2 and β‐actin antibodies (arrows). Asterisk indicates nonspecific band. Data are representative of three independent experiments.Plots showing the total dendritic length and branching points. Data presented as mean + SEM from at least three independent experiments. A total of 38 neurons were counted for each sample. Statistics were determined by the t‐test and resulted not significant.Representative example of co‐culture containing control (green), PKAN (red) human neurons, and E18 cortical mouse neurons. Control and PKAN NPCs were infected with GFP‐LV and tdT‐LV expressing vectors, respectively, and differentiated for 8 weeks.Examples of electrophysiological properties of human neurons: control individual (top); PKAN patient (bottom). Traces on the left represent trains of action potentials induced by injection of a suprathreshold current step through the patch electrode in current‐clamp mode. Middle traces show Na+ and K+ currents (down‐ and upward‐deflecting from baseline, respectively) in response to a 60 mV step from a holding voltage of −70 mV in voltage‐clamp mode. Insets on the right display enlarged portions of the traces to magnify fast Na+ currents.Histogram with percentages of recorded cells showing repetitive firing. Data presented as mean + SEM, Controls n = 32, PKAN patients n = 23 (z‐test).Data information: (A, B, and E) Scale bars 20 μm. Daniel I Orellana et al. EMBO Mol Med. 2016;8:1197-1211 © as stated in the article, figure or figure legend