Atoh1 is induced during the differentiation of human pluripotent stem cells into neurons. (A): Human induced pluripotent stem cells (iPSCs) were differentiated into neurons following the dual-SMAD inhibition protocol. By day 10 of differentiation, cells expressed neural lineage markers (PAX6 and NESTIN). By day 40 of differentiation, iPSC-derived neurons expressed neuronal markers (TUJ1 and MAP2). Cell nuclei were counterstained with DAPI. Scale bars = 50 μm. (B, C): Markers for pluripotent cells (NANOG), neural (PAX6), and neuronal (Ngn2 and NEUROD1) lineages were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) during iPSC differentiation at days 0, 10, and 20. Atoh1 expression was analyzed by qRT-PCR (B) and Western blotting (C). The data represent means ± SEM. ∗, p < .01 compared with day 0. Abbreviation: DAPI, 4′,6-diamidino-2-phenylindole.