Park BY , Hong CS , Sohail FA , Saint-Jeannet JP .

R01-DC07175 NIDCD NIH HHS , R01 DC007175-03 NIDCD NIH HHS , R01 DC007175 NIDCD NIH HHS

	Fig. 2. Expression of CXCL14 by whole-mount in situ hybridization. (A) Onset of CXCL14 expression at stage 20 in the prospective cement gland anterior to the neural plate (black arrow). Frontal view, dorsal to top. (B-D) At stage 23 CXCL14 is detected in part of the cement gland (black arrows), the dorsal anterior portion of the optic vesicle (red arrows) and the hatching gland (white arrows). (B) Lateral view, dorsal to top, anterior to right. (C) Ventral view, anterior to right. (D) Frontal view, dorsal to top. The expression of Xhe (E) and Pax3 (F) in the hatching gland (white arrows) at this stage are shown for comparison. (E-F) Frontal view, dorsal to top. (G-I) CXCL14 expression at stage 28. In addition to the cement gland (black arrows), optic vesicle (red arrows) and hatching gland (white arrows), CXCL14 is also detected in the otic vesicle (green arrows). (G) Lateral view, dorsal to top, anterior to right. (H) Dorsal view, anterior to right. (I) Ventral view, anterior to right.(J) The expression pattern of Trp2 in the pigmented epithelium of the retina is similar to CXCL14 expression in the developing eye (red arrow). Lateral view, dorsal to top, anterior to right. (K-L) Transverse section highlights the expression of CXCL14 expression in the pigmented epithelium of the retina (red arrow). Panel (L) is a higher magnification view of panel (K). CXCL14 expression is restricted to the ventral medial aspect of the otic vesicle (M). (N) At stage 35, CXCL14 expression persists in the same domains and is also detected in the head mesenchyme (yellow arrows). Lateral view, dorsal to top, anterior to right. (O-R) Transverse sections showing the expression of CXCL14 in the head mesenchyme and the cement gland (O), pigmented epithelium of the retina (O-P) and otic vesicle (Q-R). Panel (Q) corresponds to a section posterior to section shown in panel (R). For comparison at this stage Otx2 (bracket) is expressed in the ventral aspect of the otic vesicle (S) a domain that does not overlap with CXCL14 (R). di, diencephalon; hb, hindbrain; le, lens; ov, otic vesicle; re, retina; st,stomodeum.
	Fig. 4. Pax3 is necessary and sufficient for CXCL14 expression in the hatching gland. (A) Embryos injected with Pax3 morpholino (Pax3MO; 40ng) exhibit a strong reduction of CXCL14 expression in the hatching gland (arrow). In these Pax3-depleted embryos Snail2 and Xhe expression is also reduced at the neurula and tailbud stages, respectively (arrows). (B) Pax3 overexpression (0.5 ng) in the whole embryo results in a dramatic expansion of CXCL14 and Xhe expression domains at stage 23 (arrows), while Snail2 expression is reduced at stage 15 (arrow). For the embryos hybridized with CXCL14 or Xhe, anterior view is shown, dorsal to top. For Snail2 the embryos are viewed from the dorsal side anterior to top. (C) In animal explants CXCL14 and Xhe are activated by higher doses of Pax3GR mRNA as compared to Snail2. The concentrations of injected Pax3GR mRNA are 0.05 ng, 0.1 ng, 0.25 ng, 0.5 ng and 1ng. Each Real-Time RT-PCR value has been normalized to the level of EF- 1α expression.
	astl3a.1 (astacin-like metallo-endopeptidase 3 gene a1) gene expression in Xenopus laevis embryo via in situ hybridization, NF stage 23, anterior view, dorsal up. Arrow indicates hatching gland.
	dct (dopachrome tautomerase) gene expression in Xenopus laevis embryo via in situ hybridization, NF stage 28, lateral view, anterior right.
	cxcl14 (Chemokine (C-X-C motif) ligand 14) gene expression in Xenopus laevis embryo via in situ hybridization, NF stage 20, anterior view, dorsal up.
	cxcl14 (Chemokine (C-X-C motif) ligand 14) gene expression in Xenopus laevis embryo via in situ hybridization, NF stage 23, anterior view, dorsal up.
	cxcl14 (Chemokine (C-X-C motif) ligand 14) gene expression in Xenopus laevis embryo via in situ hybridization, NF stage 28, lateral view, anterior right.
	cxcl14 (chemokine (C-X-C motif) ligand 14) gene expression in Xenopus laevis embryo via in situ hybridization, NF stage 35 and 36, lateral view, anterior right
	cxcl14 (chemokine (C-X-C motif) ligand 14) is detected in part of the cement gland (black arrows), the dorsal anterior portion of the optic vesicle and the hatching gland (white arrows), in Xenopus embryo, at NF stage 23.
	cxcl14 (Chemokine (C-X-C motif) ligand 14) gene expression in Xenopus laevis embryo via in situ hybridization, NF stage 23, anterior view, dorsal up. White arrow indicates expression in the hatching gland, red arrows the dorsal aspect of the optic vesicle and black arrow the cement gland.

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