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The DNA-binding domain of Hand2 is not required for early zebrafish cardiogenesis. a, Schematic of the hand2 locus and hand2 mutants. A hand2 FLD allele was generated by targeting the 5′ and 3′ UTR sequences of hand2 (gRNA#1, gRNA#2), resulting in a deletion of 1,411 bp. A hand2 DNA-binding and phosphorylation domains-deficient allele was generated using one gRNA (gRNA#3) around the sequence encoding the DNA-binding and phosphorylation domains of Hand2, resulting in a 27-bp deletion. A hand2 dimerization domain-deficient allele was generated using one gRNA (gRNA#4) around the sequence encoding the dimerization domain of Hand2, resulting in a 3-bp deletion. b, Amino acid sequence alignment of Hand2, Hand2 Δ27 and Hand2 Δ3 generated through Clustal Omega. The three arginines in red and the threonine and serine in blue were deleted in the DNA-binding and phosphorylation domains-deficient form of Hand2 (Hand2 Δ27); the phenylalanine in green was deleted in the dimerization domain-deficient form of Hand2 (Hand2 Δ3). c, In situ hybridization showing myl7 expression in 20- and 24-hpf hand2 FLD+/+ and hand2 FLD−/− sibling embryos. d, In situ hybridization showing myl7 expression in 22- and 24-hpf hand2 Δ27+/+ and hand2 Δ27−/− sibling embryos. e, In situ hybridization showing myl7 expression in 20- and 24-hpf hand2 Δ3+/+ and hand2 Δ3−/− sibling embryos. The proportion of embryos matching the image shown is indicated in the top right corner of each image. The scale bars apply to all images.
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