Xenbase Image ID: 171108

Supplemental Figure S1. (A) Xenopus early somite stage NF14 and NF20 foregut endoderm explants contain a restricted domain of respiratory-competent cells that can respond to RA followed by Wnt/BMP pathway stimulation to activate respiratory fate. NF14 endoderm requires exogenous RA (50nM) prior to Bio (3.5uM) +BMP4 (50ng/mL) in order to express the respiratory markers nkx2-1 and sftpc in only a subset of each explant. NF20 endoderm is competent, and does not require exogenous RA in order to express respiratory markers in response to Bio+BMP4. This competence is RA-dependent, as culture of embryos in the Raldh inhibitor DEAB (10uM) prior to foregut dissection blocks the ability of Bio+BMP4 to induce the lung markers. (B-J) Gastrula anterior endoderm contributes to foregut organ lineages. Immunofluorescence analysis of optical sections through hhex:GFP transgenic X.laevis embryos. GFP expression driven by the Xenopus hhex promoter is localized to gastrula anterior endoderm (C; plane of section indicated in B) and acts as a short-term lineage label (Rankin et al., 2011). (E-I) Analysis of NF34 hhex:GFP embryos (planes of section indicated in D) reveal GFP expression is present in lateral and ventral Sox2+ pharyngeal endoderm (E,F,G), Nkx2-1+thyroid endoderm (E,E”), Sox2+ esophageal endoderm (G,H, J, J”), Nkx2-1+ respiratory endoderm (G-G”,H-H”), ventral pancreatic endoderm (I), and stomach endoderm (I, J, J”), confirming that gastrula stage AE contributes to foregut lineages. J and J” show staining of the lung/esophageal/stomach region of NF38 embryos, confirming that gastrula AE contributes to both lung bud, esophageal, and stomach endoderm. Scale bars 100uM. Abbreviations: DE, definitive endoderm; AE, anterior endoderm; PE, posterior endoderm; fb, forebrain; phe, pharyngeal endoderm; thy, thyroid; v fge, ventral foregut endoderm; d fge, dorsal foregut endoderm; sto, stomach; vp, ventral pancreas; tr, trachea; eso, esophagus; lb, lung bud; stom, stomach. Image published in: Rankin SA et al. (2018) Copyright © 2018. Image reproduced with permission of the Publisher, Elsevier B. V. Gene Synonyms Species Stage(s) Tissue sox2.S anop3, LOC108718087, mcops3, Sox-2, XLSOX-2, Xsox-2, XSox2 X. laevis Throughout NF stage 33 and 34 endoderm pharyngeal region dorsal foregut sox2.S anop3, LOC108718087, mcops3, Sox-2, XLSOX-2, Xsox-2, XSox2 X. laevis Throughout NF stage 37 and 38 esophagus endoderm nkx2-1.L nkx2.1, titf1, ttf1, Xnkx-2.1, xnkx2-1 X. laevis Throughout NF stage 33 and 34 forebrain thyroid primordium ventral foregut endoderm Image source: Published Larger Image Printer Friendly View

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