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Fig. S7
Circulation defects and pericardial edema in the nipbla/b-morphants. (A) Numbers of embryos with heart defects in nipbla/b-MOs within each class that showed circulation defects or pericardial edema at 32 hpf. (B–E) o-Dianisidine staining of developed erythrocytes. Erythrocytes were stained by incubating anesthetized living embryos in 10 mM sodium acetate, pH 4.5, 0.65% hydrogen peroxide, 40% ethanol, and 0.6 mg/ml of o-dianisidine for 15 min at 28.5°C. Control embryos (B, C) and nipbla/b-morphants with circulation defects (D, E) were stained with o-dianisidine at 32 hpf. Ventral views of the heart (B, D) and lateral views of the tail (C, E) are shown. In morphants, erythrocytes accumulated ventrally (arrow head in E) including the intermediate cell mass (ICM) (arrow in E), and yolk surface (D). (F–I) gata1 (blood cells) (F, G; 20 hpf) and fli1a (blood vessels) (H, I; 31 hpf) expression were examined by ISH in controls (F, H) and nipbla/b-morphants (G, I). Scale bars: 100 μm.