Figure 2.

GnRH-Expressing Cells Emerge after Treatment with FGF8 and DAPT

(A) Relative expression of GNRH1 at the end of the protocol in HEL11.4-derived cells (n = 4), and H9-derived cells (n = 6) treated with or without FGF8 (mean + SEM). ^∗∗p < 0.01, ^∗∗∗∗p < 0.0001 (paired ratio t test).

(B–E) Immunocytochemistry in HEL11.4-derived cells at the end of the protocol. (B) GnRH-positive neurons appeared mostly in the periphery of condensed node-like structures. These cells appeared to be interconnected by long cellular projections (a), which stained positive for MAP2 (b). (C) The GnRH-expressing cells were not proliferating, as indicated by the absence of colocalization with Ki-67 staining. (D) The GnRH-positive cells also expressed neuron-specific marker TUJ1. See also Figure S2B. (E) At the end of the protocol, the majority of the cells expressed FOXG1. Some of the GnRH-positive cells were also FOXG1 positive. All scale bars represent 50 μm.