FIG. 2.

miR-22 targets the 3′ UTR of the ERα mRNA. (A) Detailed view of the conserved target site of miR-22. The target site mutations are boxed and indicated by arrows above the sequence. Details of the mutant miR-22 variants are given below the wild-type sequence, which is in white letters on a black background. (B) miR-22 represses the luciferase activity of the wild-type full-length 3′ UTR reporter (wt) but not that of the corresponding reporter with a target site mutation (mut). The graph shows the results obtained with 293T cells cotransfected with the indicated reporter plasmids and miRNA constructs. (C) Subfragment 2 (UTR2) of the 3′ UTR of ERα contains the conserved target site of miR-22 and mediates miR-22 repression of the luciferase reporter. The graph shows the same type of experiment as in panel B. Data points are averages of triplicate samples, which were standardized to the values obtained with the control shRNA construct, which were set to 100%. Error bars represent standard deviations. (D) Q-PCR analysis of luciferase mRNA levels of the full-length 3′ UTR reporter gene in 293T cells cotransfected with the indicated miRNA/shRNA constructs. Shown are averages of triplicate samples with standard deviations.