Setdb2 is a maternally expressed gene in zebrafish and possesses H3K9me3 activity in vivo. (A) Evolutionarily conserved domain architecture of zebrafish Setdb2 protein. The Setdb2 protein is composed of a methylated DNA binding domain (MBD), a pre-SET domain, and a bifurcated SET domain. The MO1 and MO2 indicate the positions of sequences targeted by the morpholino oligonucleotides. The percentages indicate the amino acid identities between human and zebrafish Setdb2. The Arabic numbers denote the position of each domain. (B) Western blot analyses of histone methylation levels in the control and the Setdb2-deficient embryos at 10 hpf. Red and black arrows denote a reduction and an increase of the level of H3K9me3 and H3K9me1, respectively. (C) Dose-dependent reduction of H3K9me3 level in the embryos injected with indicated amounts of setdb2 morpholino. (D) Spatial and temporary expression of setdb2 during early embryonic development. The Arabic numbers at the Bottom denote the extent of methylation modification compared with histone H3 level by densitometry.
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