Fig. S5

Bioactivity of GFP sensor with 4× miR-142a-3p perfect complimentary target (PT) sites and miRNA duplex. A - Schematic of miRNA target validation GFP sensor assay. B - Schematic of miR-142a-3p perfect target (PT) sequence complementary with miR-142a-3p sequence. C - Silencing effect of miR-142a-3p on the GFP-4X PT 3′UTR gene target. Co-injection of the miR-142a-3p and GFP-4X PT 3′UTR mRNA led to suppression of GFP expression in wildtype zebrafish embryos. Expressions of the reporters were analyzed at 26 to 28 hours post fertilization. Bright-field image of embryos are shown in the upper row of each panel. DsRed mRNA was used as injection control and is shown in the lower row of each panel. In the middle row of each panel various GFP-4X PT 3′UTR sensor RNA and miR-142a-3p combinations were tested as labeled. Group of images in which embryos with co-injection of various GFP-4X PT 3′UTR mRNA and miR-142a-3p images are compared with embryos receiving respective GFP-4X PT 3′UTR sensor only. Clutch of representative embryos (7-9 embryos in each image) were photographed together in a single image to ensure valid comparison of relative green fluorescent intensity between two groups. The embryos were imaged at 2.5× magnification.