Xenbase Image ID: 154235

Fig. 5. PAPC Ser949/963 residues control protein stability and function. A, Partial sequence alignment of Xenopus laevis PAPC and human PCDH8. Conserved serine motifs at positions 949 and 963 are highlighted in red. The scheme below shows the mutant constructs in which both serines were replaced with either glutamate (S/D) or alanine (S/A). B, Immunoblot analysis of the mutated PAPCc constructs in HEK293 cells, following co-transfection with Nlk1. GFP and β-Tubulin were used as transfection and loading controls, respectively. C, ATF2 luciferase reporter assay in stage 12 Xenopus embryos injected with indicated MOs (12 ng/embryo) and mRNA (300 pg/embryo, wnt11 250 pg/embryo). The graph shows the mean±SD of three independent experiments (** p<0.01 compared with wnt11+PAPC MO injected samples). Inset: Expression analysis of PAPC constructs by immunoblotting. RLU: relative light units. D, Immunofluorescence microscopy of AC explants injected animally at the four-cell stage with mRNA for papc (I), papc-s/a (II), papc-s/d (III), or mbRFP (300 pg/embryo), and MOs (12 ng/embryo). PAPC constructs are shown in green and the plasma membrane by mbRFP in red. Nuclei were counterstained with DAPI (blue). Scale bar: 50 µm. 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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