Bantam-mediated regulation of reporter mRNAs carrying full-length D. melanogaster 3′ UTRs. (A) Schematic of reporter construct harboring the D. melanogaster hid 3′ UTR. A 2.65-kb fragment encompassing the full-length hid 3′ UTR and its natural polyadenylation signal was appended to Renilla luciferase (R-Luc-hid) and (B) tested for bantam regulation by co-transfecting the reporter construct (5 ng/well) with 100 nM 2′-O-methylated RNA oligonucleotides (anti-miRNA). (C) Other putative bantam target genes identified by TargetScan respond to anti-bantam oligonucleotide. The full-length 3′ UTRs, including their own polyadenylation signals, of D. melanogaster genes Gli, sca, and HLHmδ were fused to Renilla luciferase and tested for a specific response to anti-bantam. The TER94 3′ UTR, not predicted to have any bantam binding sites, was tested as a control. In both B and C, Renilla values were normalized to firefly values and the specific response to anti-bantam was determined by dividing Renilla values in the presence of anti-bantam by the Renilla values in the presence of control anti-lin-4 (+anti-lin-4 = 1.0).
