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Fig. 5 Regulation of dazl mRNA by miR-430 and DAZL.
(A) GFP-dazl, GFP-tdrd7 or GFP mRNA was injected with DsRed mRNA. GFP and DsRed were analyzed at 24 hpf. Insets show the gonad region. The reporter mRNA was detected by in situ hybridization with a GFP probe. (B) GFP-dazl and DsRed mRNAs were injected with or without the mRNA encoding Myc-DAZL. GFP and DsRed were analyzed at 24 hpf. Fold change of normalized GFP fluorescence relative to mock control is shown on the right. (C) Poly(A) length of GFP-dazl and GFP-dazl ΔmiR-430 site mRNAs in the presence or absence of DAZL was analyzed by RNase H-poly(A) assay at 6 hpf. (A0) shows completely deadenylated fragments by RNase H digestion with oligo (dT). The position of the RNA size marker is shown on the left. Sequences of wildtype and mutated miR-430 target sites are shown below. Nucleotides that are complementary with miR-430 seed are shown in blue. Asterisk indicates a non-specific cleavage product of the dazl 3′UTR generated with oligo (dT).