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. 2010 Jul 13;285(38):29223–29230. doi: 10.1074/jbc.M110.144576

FIGURE 2.

FIGURE 2.

Relative luminescence in plasmid constructs containing miR-320a target sites. miR-320a target regions in the 3′-UTR of AQP1 and AQP4 were identified using TargetScan and microRNA.org. These regions were cloned into luciferase reporter plasmids. Mutated 3′-UTR constructs were generated using site-directed mutagenesis. The miRNA recognition sites at cDNA corresponding to 3′-UTR (underlined) were mutated as follows: AQP1 (5′-CTGATTCCTCTCATTTAATTTGGCT-3′) and AQP4 (5′-TTGCCCCATAAGAGCAGTCGTCCGG-3′). The plasmid constructs together with anti- or pre-miR-320a were co-transfected into HeLa cells. Luciferase luminescence readings were obtained 48 h post-transfection. Relative luminescence was obtained by normalizing the values against control plasmids, pMIR-REPORTTM without any 3′-UTR insert. Statistical analyses were done using t tests. *, p < 0.05; **, p < 0.01 compared with control. The data shown are the means ± S.D., n = 3.