Fig. 1

ppp4ca and ppp4cb Spatiotemporal Expression Patterns and Their Functions on Dorsoventral Patterning in Zebrafish Embryos (A) Spatiotemporal expression pattern of ppp4ca and ppp4cb mRNA in zebrafish embryo, detected by whole-mount in situ hybridization, at indicated stages. Embryos in the first four columns were lateral views with the animal pole oriented to the top, whereas the others were oriented with anterior to the left. (B) Distribution of endogenous Ppp4c protein in zebrafish embryo at the shield stage, visualized by indirect immunofluorescence (green). The nuclei (blue) were stained with DAPI. Left: 3D reconstruction of stained embryos viewed from the animal pole by Zeiss ZEN 2009 software; right: subcellular location of Ppp4c at a higher magnification. (C) Morphological changes in ppp4ca (p4ca) and ppp4cb (p4cb) morphants at 24 hpf. Of note, p4ca or p4cb morphants exhibited variant necrosis, an off-target effect that was alleviated by coinjection of p53-MO. aMO, ppp4ca-MO; bMO, ppp4cb-MO. (D) Embryonic dorsalization induced by knockdown of ppp4ca/b was confirmed by examining several dorsoventral markers′ expressions. For the dorsal markers chd and gsc, embryos at the shield stage were dorsal views with the animal pole oriented to the top; for the ventral markers gata2 and eve1, embryos were animal-pole views at the shield stage with the dorsal side oriented toward the right. ctr, uninjected embryos as control. The ratio indicated at the right corner was the number of embryos with altered marker expression/the number of observed embryos. See also Figure S1.