Xenbase Image ID: 154231

Fig. 1. Nlk1 and PAPC interact with each other. A, Yeast two-hybrid screen to investigate PAPC and Nlk1 interaction. The intracellular domain of PAPC (PAPCc, aa715–979) and the indicated deletion constructs (white and red) were used as bait and Nlk1 (yellow) was used as prey. The interaction of bait and prey was visualized by growth on selection plates and by synthesis of β-galactosidase (blue). The last 80 amino acids of PAPC (aa900-979) are highlighted in red. B, Schematic representation of Xenopus PAPC constructs used in this study. The cytoplasmic domain (PAPCc) is drawn in red and unbound Nlk1 is emphasized by lighter appearance. C, Co-immunoprecipitation (Co-IP) of Nlk1 and PAPC in HEK293 cells. Indicated PAPC constructs were immunoprecipitated using an anti-Myc antibody and subjected to immunoblot analysis using an anti-Flag antibody to detect co-precipitated Nlk1, or anti-Myc to detect PAPC. D, Co-IP of Nlk1 and the PAPC C-terminus in HEK293 cells. PAPCc and cδ899 were precipitated using an anti-Flag antibody, and Nlk1 was detected with anti-Myc antibody. E, Immunofluorescence staining of Nlk1-Myc (green) and the indicated PAPC constructs (red) in Xenopus AC explants. Embryos were injected animally at four-cell stage with 300 pg RNA per construct. Membrane bound RFP (mbRFP) was used as control. Nuclei were counterstained with DAPI (blue). Scale bar: 50 µm. White arrowheads indicate co-localization at the plasma membrane and the asterisk marks the localization of both proteins inside the nucleus. F, papc and nlk1 were detected in Xenopus embryos by whole mount in situ hybridization. Sagittally sectioned gastrula (stage 10.5) embryos were stained with DIG-labelled antisense RNA probes (violet) for nlk1 and papc. Black arrowheads indicate position of the blastopore, while red arrowheads show the involuting mesoderm. Image published in: Kumar R et al. (2017) Copyright © 2017. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View

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