Xenbase Image ID: 147176

Figure 6. Ptk7 is required for Wnt5A-mediated papc induction (A) Four-cell embryos injected with Wnt5a mRNA (30pg per cell) in the animal pole received a second injection of Ptk7-MO (2.5ng per cell) in all animal blastomeres at the eight-cell stage. Animal caps were isolated at blastula stage, cultured for 4h at 23°C and then processed for RT-qPCR. (B) Schematic representation of Xenopus full-length PTK7 and PTK7-ICD constructs. GFP was added at the C-terminus to generate the fusions used in experiments C-E. (C) Eight-cell embryos were injected with full-length Ptk7-GFP mRNA (250pg per cell) and mRFP mRNA (50pg per cell) to reveal cell membranes, with or without Wnt5a mRNA (30pg per cell) in all animal blastomeres. Embryos were grown until early gastrula stage (st.10.5) and processed for confocal imaging. PTK7-GFP localized strictly to cell membranes in the absence of Wnt5a and partly relocalized to the nucleus in the presence of Wnt5a. (D) Eight-cell embryos were co-injected with Ptk7-ICD-GFP mRNA (300pg per cell) and mRFP mRNA (50pg per cell) in all animal blastomeres. Embryos were grown until early gastrula stage (st.10.5) and processed for confocal imaging. PTK7-ICD-GFP was found at the cell membrane and in the nucleus. (E) Eight-cell embryos were injected with the indicated amounts of PTK7-ICD-GFP mRNA, in all animal blastomeres and animal caps were isolated at blastula stage, cultured for 4h at 23°C and then processed for RT-qPCR. For all qPCR graphs, error bars represent s.e.m. values of three independent experiments with two technical duplicates. For statistical analyses, samples were compared with the respective Ctrl using unpaired Student’s t-test. Image published in: Martinez S et al. (2015) © 2015 by The American Society for Biochemistry and Molecular Biology, Inc. 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 Experiment + Assay Source Phenotypes and Disease Xla Wt + ptk7del_1-2178-GFP + animal cap explant + NF13 (RT-PCR) fig.6.a Expression Phenotype increased amount pcdh8.2.L expression in animal cap Xla Wt + ptk7del_1-2178-GFP + animal cap explant + NF13 (RT-PCR) fig.6.a Expression Phenotype increased amount pcdh8.2.L expression in animal cap Xla Wt + wnt5a + animal cap explant + NF13 (RT-PCR) fig.6.a Expression Phenotype increased amount pcdh8.2.L expression in animal cap Xla Wt + ptk7del_1-2178-GFP + NF10.5 (RT-PCR) fig.6.d Expression Phenotype mislocalised ptk7.L expression in nucleus Larger Image Printer Friendly View

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