phosphorylated smad 1 5 8  (Cell Signaling Technology Inc)

Journal: Developmental biology

Article Title: Wbp2nl has a developmental role in establishing neural and non-neural ectodermal fates

doi: 10.1016/j.ydbio.2017.06.025

Figure Lengend Snippet: (A) The ectopic expression of K81, foxd3 and zic2 are minimized by co-expression of Chd, a BMP antagonist. Red arrows point to weak posterior ectopic gene expression observed in those few embryos that were positively scored. (B) Immunostaining for phosphorylated SMAD 1/5/8, an indicator of BMP signaling when located in the nucleus. Only embryos in which ventral cells, which are subject to high levels of endogenous BMP signaling, showed nuclear staining were analyzed (b). No embryo injected only with lineage tracer (blue) showed nuclear staining in the neural plate (a), whereas a majority of embryos injected with lineage tracer plus wbp2nl mRNA showed nuclear staining (c). (C) Expression of Wbp2nl in the ventral epidermis causes ectopic expression of two neural crest genes (foxd3, zic2). Pink nuclei (lineage tracer) indicate the Wbp2nl containing cells. Ventral views, anterior to the top. (D) Ventral animal blastomeres were dissected from the 16-cell stage embryo and cultured in simple salt medium. Those dissected from uninjected control embryos never expressed foxd3 or zic2, whereas those that express Wbp2nl (red nuclei) always expressed these genes. Frequencies of the phenotypes and sample sizes (n) are given in each panel.

Article Snippet: When embryos reached stages 11–12, they were fixed and processed for whole mount immunostaining using an anti-phosphorylated SMAD 1/5/8 antibody (#9511, Cell Signaling) as previously described ( Neilson et al., 2012 ).

Techniques: Expressing, Gene Expression, Immunostaining, Staining, Injection, Cell Culture, Control