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. 2015 Aug 18;19(12):2793–2805. doi: 10.1111/jcmm.12666

Figure 3.

Figure 3

Identification of GIT1 as a target of miR‐138. (A) Diagram of the luciferase reporter constructs. Two predicted miR‐138‐targeting sequences were located in the 3′‐UTR of the GIT1 mRNA (wt‐GIT1‐1, wt‐GIT1‐2). Mutation was introduced into the target sites (underlined nucleotides) for generating mutated reporter constructs. (B) Dual luciferase reporter assay. Luciferase reporter constructs that included wide‐type or mutant GIT1 3′UTR and miR‐138 mimics or inhibitor or negative control mimics were cotransfected into 95‐D cells. 48 hrs after co‐transfection, luciferase activity was determined. (C) The expression of GIT1 mRNA in 95‐D cells transfected with miR‐138 mimics or Anti‐miR‐138 was examined by qRT‐PCR analysis, normalized to β‐actin. (D) The expression of GIT1 protein in 95‐D cells transfected with miR‐138 mimics or Anti‐miR‐138 was determined by Western blotting. β‐actin was used as an internal control. Results are presentative of three independent experiments and the error bars refer to S.D. *P < 0.05, **P < 0.01, and ***P < 0.001.