Fig. 3. CAP350 interacts with PPARs and LXR
, but not RXR
, in vitro. (A) Human PPAR
and firefly luciferase (Luc) as a control for nonspecific protein binding, and (B) PPAR
2, PPAR
, LXR
and RXR
synthesized in vitro and labeled with L-[35S]methionine were incubated with immobilized GST-KM3 or GST alone in the presence or absence of cognate ligand, as indicated. Bound radiolabeled proteins were analyzed by SDS-PAGE. The ligands used were Panagonist B for PPARs, 22-[R]-hydroxycholesterol for LXR
, and 9-cis retinoic acid for RXR
. Lanes designated Load (1/10) had 10% of the L-[35S]methionine-labeled protein added to each of the respective binding assays.