Sequential generation of neural progenitors, neurons, and astroglia in an established monolayer ES cell protocol is reproducible. A,B: Nestin expression decreases, as a proportion of total cells, from day in vitro (DIV) 14 to DIV 21. C,D: TuJ1 expression increases, as a proportion of total cells, from day 14 to day 21. E,F: GFAP expression begins by day 28. G: Quantification of nestin, TuJ1, and GFAP expression over the course of 28 days. Data are represented as mean ± s.e.m. (N = 3). Scale bar = 20 µm.