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. 2017 Sep 19;37(5):BSR20170798. doi: 10.1042/BSR20170798

Figure 4. PTEN is a downstream target gene of miR-494.

Figure 4

(A) miR-494-binding sequences in the 3′-UTR of PTEN and mutated sequence of PTEN 3′-UTR. (B) AML12 hepatocytes were cotransfected with PTEN 3′-UTR luciferase constructs (pMIR, pMIR-PTEN-wt or pMIR-PTEN-mut) and mimics NC or miR-494 mimics for 48 h. Cells were then collected and luciferase activity was determined. The level of firefly luciferase activity was normalized to Renilla luciferase activity. miR-494 suppressed the luciferase activities of the construct containing the 3′-UTR of PTEN. **, P<0.01 compared with mimics NC. (C,D) AML12 cells were transfected with mimics NC, miR-494 mimics, inhibitor NC, and miR-494 inhibitor for 48 h. The expression of PTEN mRNA and protein was detected by qRT-PCR and Western blot, respectively. All experiments were performed in triplicate, and data were represented as mean ± S.D. **, P<0.01 compared with mimics NC. ##, P<0.01 compared with mimics NC group. (E) Representative images of Western blot analysis comparing PTEN expression in liver samples in sham surgery and HIRI groups (n=6).