FIGURE 6

Validation of the function of the miR-202-3p target genes in embryogenesis. (A) Schematic illustration of rescue using shRNAs specific to the miR-202-3p target genes. Briefly, survival rates at 6 hpf were measured for wild type embryos co-injected with miR-202-3p antagomir (8 µM) and the specific shRNA (200 ng/ul) to one of the target genes at the 1-cell stage. (B) Rescue rates at 6 hpf for various shRNAs tested. More than 500 embryos were detected for each shRNA. (C) Validation of the existence of miR-202-3p binding sites in the 3′UTRs of nfkbiaa, perp and mgll genes by dual luciferase assays in HEK29T cells. Luciferase activities from constructs containing the 3′UTRs of wild type (WT) or mutated miR-202-3p binding site (MUT) for each gene were measured comparatively. NC, scrambled miR-202-3p; Agomir, synthetic miR-202-3p. Error bars, mean ± s.d., n = 3 (biological replicates).