Xenbase Image ID: 148426

Fig. 4. Effect of pou5f3.2 and pou5f3.2MO on the early development. (A) Gene expression of Xbra1 in pou5f3.2 or pou5f3.2MOinjected embryos. Synthesized RNA of GFP (a, d), pou5f3.2 (b, e) or pou5f3.2MO (c, f) was injected into lateral side of one blastomere at 2-cell stage together with b-galactosidase as a tracer, and gene expression of Xbra1 was examined at St.10.5 by whole-mount in situ hybridization. Arrowheads indicate the injection side, which is indicated by red reaction product of b-galactosidase. a–c, lateral view with the animal pole upside and the dorsal side left. d–f, vegetal view with the dorsal side up. (B) Gastrulation of pou5f3.2 or pou5f3.2MO-injected embryo. Synthesized RNA of GFP (a, e), pou5f3.2 (b, f), pou5f3.2MO (c, g) or pou5f3.2MO and pou5f3.2 (d, h) was injected into lateral side of one blastomere at 2-cell stage, together with GFP RNA as a tracer, and morphological changes were examined at St.11. a–d, vegetal view with the dorsal side up. Arrowheads indicate the injection side. e–h, GFP fluorescence showing the injection side in a, b, c and d, respectively. (C) Gastrulation defect of pou5f3.2MO-injected embryo. Effect of pou5f3.2MO on the gastrulation was evaluated as a percentage of the embryos showing gastrulation defect (black) or normal gastrulation (white) at St. 11. Bar indicates the standard deviation calculated from more than 70 samples. Gastrulation defect was caused in a dose-dependent manner, and rescued by the co-injection with synthesized RNAs of 4-base mismatch pou5f3.2. Image published in: Nishitani E et al. (2015) Copyright © 2015. Image reproduced with permission of the Publisher, John Wiley & Sons. Gene Synonyms Species Stage(s) Tissue tbxt.S bra, brachyury, ntl, t, t-a, t-b, X-bra, Xbra, Xbrachyury X. laevis Throughout NF stage 10.5 marginal zone Image source: Published Experiment + Assay Source Phenotypes and Disease Xla Wt + pou3f2 + NF10.5 (in situ hybridization) fig.4.A. b, e Anatomical Phenotype abnormally delayed closure of blastopore Expression Phenotype decreased amount tbxt.S expression in involuting marginal zone Xla Wt + pou5f3.2 MO + NF10.5 (in situ hybridization) fig.4.A. c, f Expression Phenotype decreased amount tbxt.S expression in involuting marginal zone Xla Wt + pou5f3.2 MO + NF10.5 (in situ hybridization) fig.4.A.c, f Anatomical Phenotype abnormally delayed closure of blastopore Xla Wt + pou3f2 + NF10.5 (morphology) fig.4.B. b Anatomical Phenotype abnormally delayed closure of blastopore Xla Wt + pou5f3.2 MO + NF10.5 (morphology) fig.4.B. c Anatomical Phenotype abnormal incomplete closing of the blastopore increased apoptosis in blastopore lip Larger Image Printer Friendly View

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