Fig. 5. GPC4 regulates expression of dorsal forebrain markers. Molecular analysis of neural markers in CoMo- and Gpc4Mo-injected embryos. (A-D,G-N) Frontal views; (E,F) dorsal views; (O-R) side view. Anterior is to the left. (A,B) Otx2 expression (stage 21). Arrowhead indicates forebrain expression; asterisk indicates midbrain expression. (C,D) Bf1 expression (stage 21). Arrows indicate expression in the developing telencephalon; arrowhead indicates expression in the olfactory placodes. (E,F) Expression of the posterior neural markers Krox20 (bracket) and Hoxb9 (arrowhead) in stage 21 embryos. (G,H) Fgf8 expression (stage 17). Asterisk indicates anterior neural ridge; arrowhead indicates isthmus. (I,J) Nkx2.1 expression in the ventral forebrain (stage 21); note that Nkx2.1 expression persists in GPC4-depleted embryos (J). (K,L) Emx2 expression in the dorsal forebrain of developing embryos (stage 21); note that Emx2 expression is drastically reduced in GPC4-depleted embryos (L). (M) Emx2 expression in embryos co-injected with CoMo and mouse Gpc4 (mGpc4) mRNA; note that overexpression of mouse Gpc4 does not affect Emx2 expression (compare with K). (N) Rescue of Emx2 expression in embryos co-injected with Gpc4Mo and mouse Gpc4 mRNA (compare with L). (O) Emx2 expression in a tailbud embryo injected with CoMo. (P) Emx2 expression in a tailbud embryo co-injected with CoMo and mouse Gpc4 mRNA. (Q) Loss of Emx2 expression in a tailbud embryo injected with Gpc4Mo. (R) Rescue of Emx2 expression and forehead morphology in a tailbud embryo co-injected with Gpc4Mo and mouse Gpc4 mRNA.

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