Supplementary Figure 1: Production of differentiated human motor neurons.
From: ALS-implicated protein TDP-43 sustains levels of STMN2, a mediator of motor neuron growth and repair
a, hMN differentiation, purification, and culture strategy. b, Flow-cytometric analysis of differentiated HUES3 Hb9::GFP cells. Cells not treated with the RA and SHH pathway agonist were used as negative control for the gating of GFP expression. c–f, Micrographs and quantification of purified Hb9::GFP+ cells immunostained for HB9 and counterstained with DAPI (c) (Scale bar, 10 μm) or immunostained for ISL1 and the neuronal markers β-III tubulin and MAP2 (e) (Scale bar, 20 μm). g–i, Differentiated motor neurons are electrophysiologically active as determined by whole-cell patch-clamp recordings. g, On depolarization in voltage-clamp mode, cells exhibited fast inward currents followed by slow outward currents, indicating the expression and opening of voltage-activated sodium and potassium channels, respectively. h, In current-clamp mode, depolarization elicited repetitive action potential firing. i, Response to Kainate is consistent with the expression of functional receptors for excitatory glutamatergic transmitters. Similar results were obtained for these experiments in n = 2 independent experiments.
