Figure 4.miRNAs post-transcriptionally regulate the 3'-UTR of SIRT1 mRNA. (A)
Luciferase assays were performed 24 h after transfection of the full-length
1.6 kb SIRT1 3'-UTR downstream of luciferase (SIRT1 3'-UTR)
or constructs with 4 bp in the seed-binding regions mutated (SIRT1 3'-UTR
181mt, left panel; SIRT1 3'-UTR 9mt, right panel) and control,
miR-181a, b, and c miRNA mimics (left panel) or pSuper and pSuper miR-9
expression constructs (right panel). Data are mean ± s.d. for eight
experiments. (B-C) mESCs were
transfected with individual miRNA expression constructs; protein and RNA
were isolated 48 h later. (B) Repression of SIRT1 protein was
analyzed by western blotting. Data are representative of six experiments. (C)
qRT-PCR analysis of SIRT1 mRNA levels and mature miRNA levels. Data are
mean ± s.d. for four samples.
