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jws-dvdy.21169.fig1.tif4796K Supplemental Figure 1 The expression patterns of Hr-β-catenin detected by in situ hybridization are shown in (A) the fertilized egg, and embryos at the (B) 8-cell, (C) 16-cell, (D) 32-cell, (E) 64-cell, (F) 110-cell and (G) middle tailbud stages. (B) and (G) are lateral views. (C-F) are vegetal views. As a control, embryos hybridized with sense probe are shown in (A?-G?).
jws-dvdy.21169.fig2.tif1068K Supplemental Figure 2 Specificity of antibody against Hr-β-catenin. (A) Embryos at various stages were subjected to Western blotting with antibody against Hr-β-catenin. Every lane corresponds to a sample of 20 embryos. Predicted molecular weight of Hr-β-catenin is 86 kDa. (B, C) Animal and vegetal views of 32-cell embryos. Localization of Hr-β-catenin protein in embryos injected with Hr-β-catenin MO is visualized in red. Localization to the cell boundary, but not to nuclei, is evident. (B?, C?) Merged images of Hr-β-catenin protein in red and nuclei stained green with SytoxGreen. Small green signals around the embryo in B? are test cell nuclei. The pictures were created by the maximum-intensity method, as described in the legend to Fig. 2. (D) Embryos at the 110-cell stage were subjected to Western blotting. Uninjected (WT) embryos were compared with embryos injected with Hr-β-catenin MO or Hr-β-catenin mRNA. Twenty embryos were loaded. Scale bar, 100 ?m.
jws-dvdy.21169.fig3.tif2474K Supplemental Figure 3 Specificity of antibody against Hr-Dsh. (A) Embryos at various stages and at the 110-cell stage injected with Hr-Dsh MO and mRNA were subjected to Western blotting with the antibody against Hr-Dsh. Every lane corresponds to a sample of 20 embryos. The predicted molecular weight of Hr-Dsh is 81 kDa. The Hr-Dsh band is indicated by a solid arrowhead. The band indicated by a clear arrowhead probably represents a degradation product of Hr-Dsh. (B-D?) Animal and vegetal views of embryos stained with anti-Hr-Dsh antibody and by DAB chromogenic method at the 32-cell stage, showing reduction of Hr-Dsh protein in those injected with Hr-Dsh MO, but not in those injected with control MO (C, C?). We also confirmed reduction of the signal by Hr-Dsh MO in fluorescence staining (data not shown). Scale bar, 100 ?m.

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