Fig. 3-S2

hand2-promoter-active cells were significantly decreased in yap1/wwtr1 double-knockout embryos.

(A) yap1 gene mutation by TALEN at the targeted loci. A deletion of 11 bp in the ncv117 allele results in a premature stop codon in exon 1 of yap1 (the resulting mutant Yap1 protein consists of 25 aa). Upper and lower case letters denote the target and spacer regions for the TALEN, respectively. (B) Bright field images of a yap1^wt/ncv117wwtr1^wt/ncv114 embryo (n = 4) and a yap1^ncv117wwtr1^ncv114 embryo (n = 4) at 22 hpf. Lateral view. (C) Confocal 3D-stack images (at 26 hpf) of the TgBAC(hand2:GFP) embryos with the yap1^wt/ncv117wwtr1^wt/ncv114 allele (left panels, n = 8) and the yap1^ncv117wwtr1^ncv114 allele (right panels, n = 9). Dotted lines indicate the heart region. Note that the number of hand2-promoter-active cells is decreased in yap1^ncv117wwtr1^ncv114 embryos. (D) WISH analyses of yap1^wt/ncv117wwtr1^wt/ncv114 (left panels) and yap1^ncv117wwtr1^ncv114 (right panels) embryos at 22 hpf, made using antisense probes for hand2 (upper panels, n = 4–7) and nkx2.5 (lower panels, n = 4 to 6). The confocal 3D-stack images are a set of representative images from three independent experiments.