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Fig. 1. A coiled-coil domain is necessary to localize PACSIN2 near the cell membrane in embryos. (A) Schematic representation of the wild type form of X. laevis PACSIN2 (Wt), a mutant form where a putative coiled-coil domain has been removed (∂CC) and a Wild type form of PACSIN2 with a mitochondria anchor sequence at the C-terminal end (Wt-MA). (B) Immunodetection of endogenous PACSIN2 on a sagittal section of Xenopus embryo at stage 12 using the monoclonal antibody 3D8. PACSIN2 is expressed strongly near the membrane of ectoderm (e) and mesoderm cells (m). This membrane localization is more obvious in the mesoderm and at the interface between mesoderm and ectoderm (arrowhead). (C) Embryos injected in 1 blastomere at the two-cell stage were grown to mid gastrula (12) and the over-expressed PACSIN2 detected by immunofluorescence on a sagittal section. The Wt-PACSIN2 is enriched at the vicinity of the cytoplasmic membrane while the mutant ∂CC remains cytoplasmic. yp: yolk plug; m: mesoderm; e: ectoderm. |
