Figure 3.

Rev-erbα recruits the HDAC3/NCoR corepressor complex to repress the PGC-1α gene through an intronic Rev-erb regulatory element. (A) Schematic presentation of the PGC-1α Intron1 sequence in which two conserved Rev-erbα-binding monomeric sites are closely located. (ROREd) distal RORE; (ROREp) proximal RORE. (B) Rev-erbα regulation of PGC-1α intron luciferase reporter transfected in HEK 293T cells. The control is pGL-3 promoter vector. PGC-1α luciferase reporter plasmid (0.1 μg) was used in transfection mixture along with 2 μg of pCDNA-Flag-Rev-erbα expression vector. The luciferase activities of all experiments are expressed as the mean ± SD (n = 3). (C) ChIP assay for recruitment of Rev-erbα and HDAC3 in 293T cells. (D,E) HDAC3 knockdown (D) or NCoR knockdown (E) induces endogenous PGC-1α gene expression. After shRNA or siRNA transfection, total RNA was prepared and PGC-1α gene expression was analyzed relative to GAPDH control by quantitative real-time PCR. The fold change was calculated as the relative abundance of PGC-1α mRNA in the cells receiving HDAC3 shRNA or NCoR siRNA divided by the relative abundance of PGC-1α mRNA in the cells receiving control shRNA or siRNA, which were set to 1. Results are expressed as mean ± SD. (*) P < 0.05 by paired Student's t-test.