Fig. 1.

Characterization of GMSLNs differentiated from human PSCs. a Schematic representation of differentiation protocol. b Representative image of NPCs differentiated from hESM01 in phase contrast and immunostained for neuronal markers (SOX2, PAX6, FOXP2, NCAM1, ENO2, and Nestin), counterstained with DAPI. Scale bar, 50 μm. c Representative image of neurons differentiated from iPSHD34 in phase contrast and immunostained for DARPP-32, TUBB3, GAT1, and HTT (ab109115), counterstained with DAPI. Scale bar, 50 μm. d Representative RT-PCR analysis of NPCs differentiated from hESM01 showing expression of genes: GSX2, PAX6, FOXG1, and OTX2. e Representative RT-PCR analysis of neurons differentiated from endo-iPS12 showing expression of genes: PPP1R1B, GAD1, DRD1, BCL11B, CALB1, SST, RASD2, PENK, ANO3, PDYN, GRIA1, GRIA2, GRIK2, GRIK5, GRIN1, and GRIN2B. f Representative microphotographs of neurons differentiated from hESM01 acquired via TEM showing dendritic spines and synapse formation. g Cytosolic calcium cation levels in neurons differentiated from iPSHD22 in response to depolarization of the plasma membrane with potassium chloride (KCl). Cation levels were monitored by ratiometric Fura-2 imaging. Horizontal lines on the top of the graph indicate the time of application of 2.5 mM Ca²⁺ or 130 mM KCl into the culture medium