Figure 3

Naïve mESC, differentiated to a dorsal forebrain fate, show appropriate regional and temporal specification (A) Neural differentiation protocol detailing morphogens and small molecules employed to generate dorsal forebrain progenitors and neurons from naïve PSCs. (B) Quantification of transcriptional expression of dorsal forebrain indicative genes Pax6 and Emx2 in undifferentiated (day 0, black bar) and day 7 differentiated cultures (white bar). (C,D) Representative micrographs depicting expression of Nestin, Otx2 and Pax6 in dorsal forebrain progenitors at day7. (E) Maturing dorsal forebrain neurons, expressing Tuj1, show transient Tbr2 expression at day 11, and mature dorsal forebrain markers inclusive of (F) Tbr1 and (G) Ctip2 by day 14. Images show culture overviews, while inserts show immunocytochemical labeling at the resolution of individual cells. Data (n = 4) represented as mean ± SEM, Students t-test. Scale bar = 100 um. ***p < 0.001.