Fig. 4.

ES cell-derived Rx⁺ progenitors differentiate into early precursor neurons of the dorsal and ventral hypothalamus. (A–C) E13.5 mouse hypothalamus. Coronal (A) and parasagittal (B and C) sections were stained with DAPI and antibodies against Otp (A and C; dorsal domain, d), SF1 (A and C; ventral domain, v), and Foxb1 (C; mb). III, third ventricle; lv, lateral ventricle; or, optic recess; and p, pituitary. (C) Immunostaining within the square (hypothalamic region) in panel B. (D) Schematic of reaggregation culture of FACS-sorted Rax–GFP⁺ progenitors. (E–J) The aggregates were cultured in DMEM/F12/KSR medium with (F, G, I, J, and L) or without (E, G, H, J, and K) 30 nM Shh (days 7–13). Frozen sections were immunostained for Otp (E–G, K, and L), SF1 (H, I, and J), Tuj (E and F), Nkx2.1 (H and I), and Brn2 (K and L). (G and J) Statistical significance was examined by Mann–Whitney's test. ***, P < 0.001.