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. 2008 Aug 28;105(36):13421–13426. doi: 10.1073/pnas.0801613105

Fig. 3.

Fig. 3.

MiR-34a binding site within SIRT1 3′ UTR mediates miR-34a translational repression. (A Top) miR-34a and the miR-34a-binding site in the 3′ UTR of SIRT1. (Bottom) Design of a miR-34a reporter vector containing a CMV promoter driving expression of a luciferase cDNA fused to the SIRT1 3′ UTR or to a mutated SIRT1 3′ UTR. (B) The 3′ UTR of SIRT1 mediates miR-34 control of SIRT1 expression. WT HCT116 cells were transfected with a reporter vector consisting of a luciferase cDNA fused to the 3′ UTR of SIRT1 which contains a binding site of miR-34. Another vector contained the luciferase cDNA fused to a SIRT1 3′ UTR with a mutant miR-34a-binding site. The cells were also transfected with a CMV-Renilla luciferase vector as an internal standard. PremiR-34a decreases expression of luciferase containing a WT miR-34a binding site (Left) but not a mutant binding site (Right). (C) miR-34 does not change SIRT1 mRNA. HCT116 cells were transfected with antisense-miR-34a or PremiR-34a, and total RNA was harvested and analyzed for SIRT1 by Northern blotting.