Regulation of constructs containing putative target sites that rely on complementary pairing with nucleotides 3–9 of bantam. (A) Constructs with synthetic 3′ UTRs harboring binding sites with complementarity to either nucleotides 2–8 or 3–9 of bantam are repressed to similar degrees. R-Luc-synthetic(2–8), R-Luc-synthetic(3–9), or R-Luc-flipped reporter plasmids were transfected (5 ng/well) along with F-Luc as the transfection control in D. melanogaster S2 cells. Both R-Luc-synthetic(2–8) and R-Luc-synthetic(3–9) constructs were repressed to similar extents relative to control R-Luc-flipped. (B) Full-length 3′ UTRs of three candidate genes, nct, I(3)73Ah, and CG34131, were individually appended to Renilla luciferase and tested for bantam repression by co-transfecting with either anti-bantam oligonucleotide or control anti-lin-4. Normalized Renilla values in the presence of anti-bantam were divided by values in the presence of control anti-lin-4 (+anti-lin-4 = 1.0).
