Control of Gene Expression by the Retinoic Acid-Related Orphan Receptor Alpha in HepG2 Human Hepatoma Cells
Figure 5
qPCR analysis of immunoprecipitated chromatin shows that RORα binds to the regulatory regions of SPARC, PLG, G6PC, NR1D2 and AGRP in HepG2 cells.
Chromatin was prepared from stably-transfected HepG2 cells over-producing RORα1 (A) or not (B) and fragmented by sonication. Chromatin fragments were then immunoprecipitated with a specific anti-RORα antibody (black bars) or with a nonspecific IgG as a negative control (white bars). qPCR (normalized to input) was used to amplify genomic fragments covering the ROREs identified in silico in the SPARC, PLG, G6PC, NR1D2 (intron 5) and AGRP genes. qPCR amplification of a DNA region located in the SPARC gene but devoid of RORE was used as a negative control (SPARCn). Data are represented as the means +/− S.E.M. (n = 3). *: P<0.05, **: P<0.01 between samples treated with the anti-RORα antibody and samples treated with nonspecific IgG.