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. 2015 Mar 29;6(16):14153–14164. doi: 10.18632/oncotarget.3686

Figure 6. miR-221/222 targets TIMP-2.

Figure 6

(A) Software prediction of miR-221/222 determines a potential binding site in the TIMP-2 3′ UTR. The nucleotide sequence illustrates the predicted base pairing between miR-221/222 and the TIMP-2 3′ UTR. (B) In vitro luciferase reporter assay in HEK293 cells. miR-221/222 mimics or inhibitors and their negative controls were co-transfected with reporter plasmids containing the wild-type (wt) TIMP-2 3′UTR and mutant (mu) TIMP-2 3′UTR (*p < 0.05). (C) Cells were transfected with miR-221/222 mimics or inhibitors, and TIMP-2 protein levels were detected by western blot assays. GAPDH protein was assayed as a control. (D) siTIMP-2 could inhibit the expression of TIMP-2. (E) Panc-1 cells were transfected with siTIMP2; protein levels of MMP-2 and MMP-9 were detected by western blot assays. (F and G) Inhibition of TIMP-2 by siTIMP2 mimics miR-221/222-induced invasion. (*p < 0.05).