Figure 5.

PPARs bind to the PPRE in the -2366/-2339 region of the human adrp gene. A, B, C and D, EMSAs were performed with ³²P-labelled either ADRP or mutated ADRP (ADRPmut) oligonucleotides in the presence of purified PPARα (A), purified PPARδ (B), in vitro transcribed/translated PPARγ1 (C), purified PPARγ2 (D) and/or purified RXRα proteins. Supershift experiments were carried out using anti-PPARα (A), anti-PPARδ (B) or anti-PPARγ (C and D) antibodies. Unlabelled oligonucleotides (ADRP, ADRPmut or ACO) were used at 10- or 100-fold molar excess to the labelled probe to perform competition assays. Closed and open arrowheads indicate the specific bands and the supershift bands, respectively.