Figure 5

Association of genomic regions of PPAR target genes with PPARs and their partner proteins. Chromatin was extracted from HEK293 cells that had been treated with solvent (DMSO) or for 120 minutes with 100 nM GW7647. The association of PPARα, RXRα and pPol II was monitored by ChIP assays with respective antibodies on genomic regions of the eight PPAR target genes that are (a) close to the TSS, (b) upstream of the TSS and (c) downstream of the TSS; for location see Figure 3 and Table 2. Since the APOC3 gene is not expressed in HEK293 cells, the data for its four genomic regions were obtained using chromatin derived from HepG2 cells. Real-time quantitative PCR was performed on chromatin templates and the fold change of the antibody-precipitated template in relation to an IgG-precipitated specificity control template was calculated. (d) PPARα shows specific association with 15 of the 23 tested regions and the relative association with these regions is shown. Columns represent means of at least three experiments and bars indicate standard deviations. Two-tailed Student's t-tests were performed to determine the significance of association in reference to IgG controls (*p < 0.05, **p < 0.01, ***p < 0.001).