Figure 2.

TCF12 and Fli-1 are directly regulated by miR-200 s. (a) Luciferase activity of CAFs co-transfected with different ratios of the indicated miR-200 s or control vector. TCF12 and Fli-1 reporters driven by wild-type 3'-UTRs (left panel) and TCF12 and Fli-1 reporters driven by 3'-UTRs with wild-type or mutated miR-200 s-binding sites (right panel) are shown. miR-Ctrl, non-miRNA-expressing control; wt UTR, wild-type 3′-UTR of TCF12 or Fli-1; mut UTR, mutant-binding sites for miR-200 s in the target mRNA. All the luciferase reporter assay results were normalized to Renilla luciferase, and the data are presented as the mean±S.D. (n=3; *P<0.05, ANOVA followed by the Student–Newman–Keuls test). (b) Lipofectamine 2000 was used to transiently transfect the miR-200 s mimics into CAFs or siRNA into NFs. Fli-1 and TCF12 expression was examined by qRT-PCR. β-Actin was used as an internal control. The data are presented as the mean±S.D. (n=3; *P<0.05, Student's t-test). (c and d) Western blot analysis of endogenous Fli-1 (c) and TCF12 (d) expression in the indicated fibroblasts. β-Actin was used as a loading control