Figure 2
MiR-194 directly targets COUP-TFII mRNA. (a, b) C3H10T1/2 cells were transfected with miR-194 precursors or anti-miR-194. Expression levels of miR-194 and COUP-TFII mRNA were examined using qRT-PCR (n=3). Values represent mean±S.D. *, P<0.05 and **, P<0.01 compared with miR-CTL. (c) C3H10T1/2 cells were transfected with miR-194 precursors or anti-miR-194, and Western blotting analysis was performed. The data were obtained from three independent experiments. (d) Computational analysis of one putative complementary sequence for miR-194 in the 3′-UTR fragment of COUP-TFII. The wild type (WT) or mutant type (MT) construct was inserted into pmirGLO reporter vector. (e) The pmirGLO, pmirGLO COUP-TFII-3′-UTR-WT (WT) and pmirGLO COUP-TFII-3′-UTR-MT (MT) vectors were co-transfected with miR-CTL and miR-194 or anti-miR-194, respectively, into C3H10T1/2 cells. Luciferase activities were measured from the cell lysates after 24 h. Relative renilla luciferase activity was normalized to that of firefly luciferase. The values were normalized by the miR-CTL-treated group (n=3). **, P<0.01. All experiments were independently repeated at least three times
