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. 2014 Sep 3;34(11):1826–1836. doi: 10.1038/jcbfm.2014.152

Figure 4.

Figure 4

MicroRNA-181a (MiR-181a) inhibits the expression of Kruppel-like factor 6 (KLF6) by targeting its 3′-untranslated region (3′-UTR). (A) Quantitative RT-PCR analysis of KLF6 in glioma endothelial cells (GECs) with the expression of miR-181a changed. (B) Western blot analysis of KLF6 with the expression of miR-181a changed. Representative protein expression and their integrated light density values (IDVs) are shown. Data represent means±s.d. (n=5, each). *P<0.05 versus miR-181a (+)-NC group, #P<0.05 versus miR-181a (−)-NC group. (C) Schematic representation of the predicted binding sites for miR-181a in the 3′-UTR of KLF6 (KLF6-3′UTR-Wt) and the site mutagenesis design for the reporter assay (KLF6-3′UTR-Mut1, KLF6-3′UTR-Mut2, and KLF6-3′UTR-Mut3). Luciferase reporter assay of human embryonic kidney (HEK) 293T cells cotransfected with the reporter plasmid (or the corresponding mutant reporter) and the indicated miRNAs. Renilla/firefly luciferase ratios were calculated and further normalized to the without miRNA group which was set as 1. Data represent means±s.d. (n=5, each). *P<0.05 versus pre-miR-181a-NC group. NC, negative control.