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. 2017 Aug 3;38(4):2155–2165. doi: 10.3892/or.2017.5875

Figure 6.

Figure 6.

miR-195 directly targets DCUN1D1. (A) Sequence alignment of miR-195 and seed match region in the 3′-UTR of DCUN1D1 as well as the mutated 3′-UTR of DCUN1D1 are shown. (B) The 3′-UTR sequence of DCUN1D1 was amplified and subcloned into the pmirGLO luciferase reporter vector. HEK293T cells were co-transfected with wild-type (WT) or mutant (Mut) 3′UTR vectors and miR-195 mimics using Lipofectamine 2000. After 48 h, the cells were assayed for luciferase activity using the Dual-Luciferase Reporter Assay System. The firefly luciferase activities were normalized to Renilla luciferase activity. (C) Quantitative real-time PCR was used to evaluate the DCUN1D1 mRNA expression in TU-177 cells transfected with miR-195 or control. (D and E) Western blot was used to evaluate the protein expression of DCUN1D in TU-177 cells transfected with miR-195 or control. Data are expressed as the mean value ± SD. **P<0.01. (F) DCUN1D1 mRNA was inversely associated with miR-195 in 122 pairs of LSCC tissues using linear regression models.