Fig. 3

(a–d) chd mRNA expression detected by in situ hybridization in WT embryos at indicated stages. The shown were animal-pole views with dorsal to the right. (e–h) Chd protein distribution in WT embryos at different stages. Embryos were immunostained using an anti-Chd antibody and observed by confocal microscopy. Representative z-stack images were shown with dorsal to the right. The colour bar blow each picture illustrated signal level changes from ventral to dorsal with the ratio of Chd/DAPI at the two extremes indicated (also see Supplementary Fig. 2d). (i–k) Confocal images of Chd in different regions of embryos at the shield stages. Different regions boxed in g were viewed in an optical equatorial plane within the blastodermal margin. Note the presence of Chd in the ventralmost region (i). (l) Subcellular localization of Chd in embryos. A region boxed in g was enlarged. (m–p) Relationship between chd mRNA and Chd protein locations in embryos at the shield stage. Embryos were probed with antisense chd RNA probe and then immunostained using anti-Chd and anti-β-catenin antibodies along with DAPI staining. A representative embryo was observed under a confocal microscope in bright field (m) or in fluorescent field (n,o) with a focus plane at the blastodermal margin. In p, the confocal fluorescence image in n was superimposed on the bright/dark image in m. All images were orientated with dorsal to the right. The border of the chd mRNA area was labelled with a circle.