Figure 3

SIRT1 is a new target for regulation by miR-204. (A) illustration of SIRT1 3’UTR as well as the seed sequence of miR-204 showing the computationally predicted target region on the 3’UTR of SIRT1 mRNA. (B) miR-204 decreases SIRT1 3’UTR luciferase activity. HEK293T cells were transfected with 2μg of pGL3 luciferase vector containing either SIRT1 3’UTR or SIRT1 MUT 3’UTR (with an A-to-G mutation in miRNA-responsive element). Cells were co-transfected along with 50 nM miR-204 precursor for 24 h and then lysed and assessed for luciferase activity. Mean-S.E. (n =3). (C) SIRT1 mRNA levels in AGS and BGC transfected with 50 nM miR-204 precursor for 72 h. Mean-S.E. (n=3). (D) Western blotting of SIRT1 protein levels. SIRT1 levels in AGS and BGC transfected with 50 nM miR-204 precursor for 48 h (n=3). **P < 0.01. *P < 0.05.