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Fig. 1 Ventral gsc overexpression induces complete secondary axes. (A-I) Live zebrafish embryo injected with gfp (A-C) or gsc (D-I) RNA. (A-F,H,I) DIC and fluorescence merge; (G) DIC. (A,D) Shield (arrowhead), animal view. (B,F) Bud; arrowheads indicate notochord, asterisk indicates prechordal plate. (C) 1 dpf, side view. (E) Bud; side view, secondary prechordal plate (asterisk). (G-K) 1 dpf, dorsal view. (J,K) dlx2a and shha (red). (J) gfp RNA injected. (K) gsc RNA injected showing duplicated dlx2a expression (brackets): staining differs from the control because the embryo had a ventral third eye. (L) Schematic of injection procedure, clones in green; one clone was generated per embryo. dien, diencephalon; e, eye; fp, floor plate; hg, hatching gland; myo, myotomes; noto, notochord; tele, telencephalon.