Fig. S1

Efficiency test of setdb2 and dvr1 morpholinos. (A) Knockdown of Setdb2 protein by specific morpholino in vivo. Western blot analysis of embryos injected with setdb2 control or setdb2 morpholinos at the indicated dos. Every control and morpholino sample had one repeat as shown. The proteins extracted from 20 embryos at 24 hpf were loaded into each lane. Histone H3 was used as a loading control. (B) Knockdown efficiency of dvr1 mo^ATG. Schematic presentation of the reporter plasmid used to test the efficiency of morpholino knockdown. The full-length 5′UTR and immediate 159-nt downstream of translational start codon (ATG) of dvr1 were fused to the ATG-deleted open reading frame of EGFP in the pCS2+ plasmid. The red bar denoted the targeted site recognized by morpholino. Co-injection of EGFP reporter mRNA with either control or dvr1 mo^ATG into wild-type embryos. Fluorescent stereomicroscope showed EGFP expression at 8 hpf. Co-injection of mcherry mRNAs was used as a control. (C) Knockdown efficiency of dvr1 mo^SB. Schematic representation of dvr1 mo^SB target site. Quantitative real-time RT-PCR analysis of dvr1 mRNA level in the embryos injected with either control or 16 ng dvr1 mo^SB at different developmental stages. Results were presented as means ± SD from three independent experiments. Zebrafish housekeeping gene gapdh was used as an internal control for data normalization. Asterisks indicate statistically significant differences (^***P<0.001).