Figure 1.

Neuroectoderm Generated with Both EB and AD Paradigms Holds Normal Patterning Potency along the A-P and D-V Axes

(A–D) Without morphogen treatment, human neuroectoderm cells will take an anterior-dorsal fate as demonstrated by uniform PAX6, SOX1, and FOXG1 expression in committed regional neuroepithelia under both EB (day 17) (A) and AD (day 12) (C) conditions. Insets show Hoechst counterstaining of nuclei. Scale bars, 50 μm. Graphs (B) and (D) show the percentage of positive cells in (A) and (C), respectively. Data are presented as mean ± SEM of three independent experiments.

(E–G) Confocal images show NKX2.1⁺/PAX6⁻ ventral progenitors yielded from both EB (E) and AD (F) differentiation paradigms. Insets show Hoechst counterstaining of nuclei. Scale bars, 50 μm. Graph (G) shows the percentage of positive cells in (E) and (F). Data are presented as mean ± SEM of three independent experiments.

(H and I) SHH treatment at days 10–17 in EB (H) or days 4–12 in AD (I) differentiated cells results in efficient ventralization as shown by increased NKX2.1, while PAX6 expression is decreased. Data are presented as mean ± SEM of three independent experiments. Unpaired two-tailed Student's t test: ^∗p < 0.05, ^∗∗p < 0.01.

(J and K) RA treatment at days 10–17 in EB (J) or days 4–12 in AD (K) differentiated cells results in efficient caudalization as shown by increased HOXB4, while FOXG1 expression is decreased. Data are presented as mean ± SEM of three independent experiments. Unpaired two-tailed Student's t test: ^∗∗p < 0.01.

See also Figure S1.