Fig. 3.

ES cell-derived Rax⁺ progenitors differentiate into early neuronal precursors of the dorsal and ventral hypothalamus. (A–C) Immunohistochemical analysis of the E10.5 mouse brain with Nkx2.1 (A–C), Pax6 (A and B) and Rax (B and C) antibodies. Coronal (A and B) and parasagittal (C). dh, vh, rh, and ch, dorsal, ventral, rostral, and caudal portions of the hypothalamus; ge, ganglionic eminence. The caudal hypothalamus (its ventral portion) is Nkx2.1⁺ and Rax⁻. (D–I) Subregional marker expression in SFEBq/gfCDM-cultured ES cells on day 7. Frozen sections of ES cell aggregates treated with 30 nM Shh (D, E, G, and H) or 10 μM cyclopamine (D, F, G, and I) were immunostained for Rax and Nkx2.1 (D and F) or Pax6 (G–I). (D and G) Statistical significance vs. the control (gfCDM) was evaluated by the Dunnette test. *, P < 0.05; **, P < 0.01; ns, not significant. (J and K) Effects of Shh (30 nM) and cyclopamine (10 μM) on the percentages of Rax–GFP⁺ (J) and Sox1–GFP⁺ (K) cells in SFEBq/gfCDM culture.