Extended Data Fig. 6. Transient inhibition of EZH2 triggers enhanced neuronal maturation even in the presence of astrocytes.
a, Transient inhibition of EZH2 at NPC stage does not trigger the generation of astrocytes under synchronized neurogenesis conditions. Representative images of MAP2 and GFAP staining at d40 and d65 in cultures derived following the synchronized neurogenesis paradigm upon EZH2i versus DMSO. Primary rat astrocytes are positive control for the GFAP staining. b, Schematic of experimental paradigm for the Ca2+ imaging of hPSC-derived neurons in EZH2i and DMSO conditions in co-culture with primary rat astrocytes. hPSC-derived neurons were infected with GCamp6m lentiviruses four days before dissociation and prior to seeding onto rat primary astrocytes. c, Representative images of hPSC-derived neurons expressing GCamp6m in co-culture with rat primary astrocytes. d, Amplitude and frequency of spontaneous Ca2+ spikes of individual neurons. DMSO + Astro n = 225, EZH2i + Astro n = 264 (neurons from 2 independent experiments). e, Synchronicity rate of spontaneous network activity in EZH2i versus DMSO conditions in co-culture with rat astrocytes (n = 6 FOV from 2 independent experiments). Two-tailed Welch’s test (d, e) Scale bars are 100 μm. FOV, Field of View.