Fig. 5. Ezrin2 regulation analyzed by 2D gels, western blotting and immunostaining of sections. (A) Close-up view of three out of seven gels (Cy3 and Cy5 labeled) in the region of the ezrin2 spot (circled red). (B) Quantification of ezrin2 spot intensities of the DIGE 2D gel experiment. (C) Western blot analysis of wild-type, mz-oep (ectoderm) and cyclops mRNA injected (mesendoderm) embryos at 8 hpf (corresponding to 80% epiboly stage in wild-type embryos). After analysis with phospho-ERM (p-ERM) antibody, blots were stripped and re-probed with ERM antibody. The lower part of the blot was probed for tubulin as a loading control. (D) Wild-type embryos were sectioned at the shield stage (6 hpf; sagittal section through the shield region) and at 80% epiboly (8 hpf; transverse section through the emerging body axis, paraxial region) and stained with phospho-ERM (p-ERM) antibody. In mesendodermal cells the level of phosphorylated ERM proteins is increased. Insets show section planes.