Fig. S3
miR-153 targets snap-25b. (A) GFP reporter constructs were created by fusing the reading frame of GFP to the snap-25b 32UTR. Three predicted miRNA recognition elements (MREs) were identified in the snap-25b 32 UTR. The miR-153 sequence is indicated in red and the corresponding snap-25a UTR sequence is shown in green. (B) Single cell zebrafish embryos were injected with mRNAs derived from GFP reporters lacking a UTR (GFP), fused to the full length snap-25b UTR (GFP+snap-25b), or mutant version of the snap-25b UTR lacking all MREs (GFP+snap-25bΔMRE1, 2&3). Embryos were injected in the presence or absence of exogenous miR-153 or morpholinos against miR-153 (miR-153MO). Fluorescence levels were examined at 1 dpf. Clusters of embryos (~30) are shown. (C)Lysates from ~100 embryos were prepared from embryos treated as in B and GFP protein levels were determined by western blotting using antibodies against GFP or control antibodies against α-tubulin.