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Fig.1. Effects of C-cad-MO. (A–H) Late gastrula sagittal sections, after dorsovegetal injection of control MO or C-cad-MO (40 ng/embryo). (A–D) In situ hybridization. gsc-expressing anterior mesoderm (arrowheads, A,B, dark purple); Xbra-expressing chordamesoderm (arrowheads, C,D). Injected cells, magenta. (E–H) High magnification, injected mesoderm cells (green) counterstained for actin (red). Brachet's cleft (arrows) separates mesoderm (left) and ectoderm (right). Tightly packed cells in control MO (E) and C-cad-MO (F) anterior mesoderm. Chordamesoderm cells are spindle-shaped in controls (arrowheads, G), but rounded in C-cad-MO embryos (arrowheads, H). (I) RT-PCR for cadherins and protocadherins in embryos injected with C-cad-MO (40 ng) or C-cadherin mRNA (2 ng). (J,K) RT-PCR to show gsc and Xbra expression after C-cad-MO or C-cadherin mRNA (J) or β-catenin MO (K; 6 ng/embryo) injection. (L–L′) β-catenin membrane localization is reduced by C-cad-MO (10 ng) in anterior mesoderm. β-catenin staining (L′, red, arrowheads) in C-cad-MO cells co-injected with GFP mRNA (L, green; L′, merged). Anterior mesoderm is to the left of Brachet's cleft (arrows). Scale bars: 50 μm. |
