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. 2008 Aug 28;27(19):2616–2627. doi: 10.1038/emboj.2008.172

Figure 3.

Figure 3

miR-324-5p modulates Gli1 expression and function through binding to the 3′UTR sequence. (A) Schematic representation of 3′UTR sequence from human Gli1 and the corresponding luciferase reporter vector, indicating the potential miRNA-binding sites on the sequence. (B) miRNA levels (evaluated by Q-PCR, relative to RNU6B and RNU66 housekeeping controls) in Daoy cells 24 h after transfection with mimic negative control (basal) or the indicated miRNAs (overexpressed). (C) Levels of luciferase activity in Daoy cells overexpressing the indicated miRNAs and transfected with the Gli1 wild-type 3′UTR vector. Data are indicated as ratios with respect to pGL4 control vector-transfected cells (ctr). *P<0.05. (D) Levels of luciferase activity in Daoy cells overexpressing miR-324-5p and transfected with the Gli1 wild-type 3′UTR vector (wt) or its mutant derivative lacking the miRNA-binding sites (mut). Data are indicated as ratios with respect to pGL4 control vector-transfected cells (ctr). *P<0.05. (E) Western blot analysis of endogenous Gli1 protein levels in Daoy cells overexpressing miR-324-5p or miRNA mimic negative control (ctr). All data in (B–D) are average values±s.d. from at least three experiments, each carried out in triplicate.