Extended Data Fig. 3 | Characterization of converted neurons from mouse
and human astrocytes. a, b, Conversion of mouse and human astrocytes to
neurons. Cells were immunostained with the indicated markers after
conversion from mouse cortical astrocytes (a) or human astrocytes (b).
Converted glutamatergic (marked by VGlut1) and GABAergic (marked by
GAD67) neurons constituted approximately 90% and 80% of total TUJ1-marked
neurons from mouse and human astrocytes, respectively. Data were based on
4 (a) or 5 (b) biological repeats and represented as mean ± s.e.m. Scale bars,
30 μm (a); 40 μm (b). c, d, Efficient conversion from human astrocytes to
neurons. Converted neurons were characterized by immunostaining with TUJ1
and MAP2 (c). Scale bar, 80 μm. n = 4 biological repeats. d, These neurons are
functional as indicated by repetitive action potentials (top left), large currents
of voltage-dependent sodium and potassium channels (top right) and
spontaneous postsynaptic currents after co-culture with rat astrocytes
(bottom). Indicated in each panel is the number of cells that showed the
recorded activity versus the number of cells examined. e–h, Electrophysiological
characterization of neurons converted from mouse (e) and human (f)
astrocytes, showing spontaneous excitatory and inhibitory postsynaptic
currents that could be sequentially blocked with the inhibitors against the
excitatory (NBQX and APV) and inhibitory (PiTX) receptors, indicative of their
secretion of glutamine and GABA neurotransmitters. g, h, Control shRNA
(shCtrl)-treated mouse (g) and human (h) astrocytes failed to show action
potentials (top), currents of voltage-dependent channels (middle) or
postsynaptic events (bottom). The number of cells that showed the recorded
activity versus the total number of cells examined is indicated on the top right
of each panel.