|
Fig. S3 GFP sensor target validation assay for vegfab 3′UTR. A - Schematic of GFP-vegabf 3′UTR transcript containing two miR-142a-3p predicted binding site (site “A”, blue; site “B”, purple). Predicted seed complementarity sequences are boxed in red colour. B - Silencing effect of miR-142a-3p on the GFP-vegfab 32UTR gene target. Co-injection of the GFP-vegfab 3′UTR mRNA and miR-142a-3p led to moderate suppression of GFP expression in wildtype zebrafish embryos. Expressions of the reporters were analyzed at 26 to 28 hours post fertilization. DsRed mRNA was used as injection controls and is shown in the lower panel. Upper panel displays GFP-vegfab 3′UTR sensor RNA and miR-142a-3p combinations. Group images of embryos that were co-injected of various GFP-vegfab 3′UTR mRNA and miR-142a-3p are compared with embryos receiving respective GFP-vegfab 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.