Figure 3

TIMP3 is a target for miR-221 and miR-222. (A) Schematic representation of the putative binding sites in the TIMP3 mRNA 3' UTR for miR-221 and miR-222 (the seed sequence (AGCUACAU) was identical, as shown). (B) Cells were transfected with As-miR-221/222 or miR-221/222 and TIMP3 protein levels were detected by western blot assays. β-actin protein was assayed as a control. (C) pGL3-TIMP3-3'UTR luciferase constructs were transfected into cells that were then treated with As-miR-221 and/or As-miR-222. Luciferase activity was determined 48 h after transfection. The ratio of normalized sensor to control luciferase activity is shown. Error bars represent standard deviation and were obtained from three independent experiments. * P <0.05 compared with control group, ** P <0.01 compared with control group.