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Figure 3—figure supplement 1. Validation of Bcat-MO embryos. (A) Immunostaining of Tg(WntREs:dEGFP)Vlem Xenopus tropicalis gastrula embryos show the loss of nuclear Bcat and GFP expression from the Wnt-reporter transgene in Bcat-MO embryos demonstrate effective knockdown. (B–C) Co-injection of RNA encoding a human S37A-stabilized Bcat recues the Bcat-MO-ventralized phenotype. In situ hybridization of gastrula (B) and tailbud embryos (C). In Bcat-MO gastrula, fst and admp are downregulated in the dorsal organizer, whereas wnt8a and ventx3.1 are upregulated consistent with a ventralized phenotype. (C) In situ hybridization of markers for the liver (nr1h5), stomach, lung, intestine and pharynx (osr1 and islet1) show that co-injection of human S37A-stabilized Bcat RNA rescues the ventralized phenotype and disrupted gut development in Bcat-MO tailbud embryos. Image published in: Mukherjee S et al. (2020) © 2020, Mukherjee et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Gene Synonyms Species Stage(s) Tissue fst foll, follistatin, xfs X. tropicalis Throughout NF stage 10.5 involuting marginal zone involuted dorsal mesoderm admp X. tropicalis Throughout NF stage 10.5 endoderm dorsal endomesoderm involuted dorsal mesoderm axial mesoderm ventx3 vent-3, ventx3.1 X. tropicalis Throughout NF stage 10.5 ventral endoderm ventro-lateral marginal zone wnt8a wnt-8, wnt8, Xwnt-8, Xwnt8 X. tropicalis Throughout NF stage 10.5 ventro-lateral marginal zone involuted ventral mesoderm nr1h5 for1 X. tropicalis Throughout NF stage 35 and 36 liver osr1 odd1 X. tropicalis Throughout NF stage 35 and 36 lung stomach cement gland isl1 islet-1, islet1, Xisl-1, Xislet-1 X. tropicalis Throughout NF stage 35 and 36 pharynx stomach intestine spinal cord pharyngeal region eye profundus ganglion midbrain Image source: Published Experiment + Assay Source Phenotypes and Disease Xtr Wt + ctnnb1 MO + NF10.5 (immunohistochemistry) Fig.3.S1.A Anatomical Phenotype abnormal Wnt signaling pathway Expression Phenotype decreased amount ctnnb1 expression in nucleus Xtr Wt + ctnnb1 MO + NF10.5 (in situ hybridization) Fig.3.S1.B Expression Phenotype decreased amount admp expression in axial mesoderm decreased amount admp expression in dorsal endomesoderm decreased amount admp expression in endoderm absent fst expression in dorsal involuting marginal zone absent fst expression in involuted dorsal mesoderm increased amount ventx3.1 expression in animal cap increased amount ventx3.1 expression in blastocoel floor increased amount ventx3.1 expression in endoderm increased amount wnt8a expression in blastocoel floor increased amount wnt8a expression in ectoderm increased amount wnt8a expression in endoderm increased amount wnt8a expression in the animal cap increased amount wnt8a expression in ventral marginal zone Xtr Wt + ctnnb1 MO + NF35/36 (in situ hybridization) Fig.3.S1.C Anatomical Phenotype wholly ventralized embryo Expression Phenotype absent isl1 expression in embryo absent nr1h5 expression in embryo absent osr1 expression in embryo Larger Image Printer Friendly View

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