Figure 4.

miR-29b targets multiple members of the BH3-only family. (A) Schematic representation of predicted miR-29b-binding sites in the 3′UTRs of multiple mouse BH3-only mRNAs. Nucleotides of the 3′UTR containing miR-29b-binding sites are listed in parentheses. (B) Sequence and alignment of the miR-29b-binding sites in the 3′UTRs of multiple BH3-only mRNAs. The predicted base-pairing of miR-29b with target recognition seed sequence is shown in bold. (C) Luciferase activity was measured 48 h after transfection of HEK293T cells with reporter plasmids in which regions of either wild-type (wt) or mutant (mut) 3′UTRs of genes listed in A were each fused downstream from the firefly luciferase gene. Reporter plasmids were transfected either alone (vector), together with 20 nM miR-29b (miR-29b), or together with 20 nM cel-miR-67 (ctrl). Expression was normalized by taking the ratio of firefly to renilla luciferase, and is plotted relative to vector alone. Data are mean ± SEM of at least three independent experiments.