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Fig. 3. CAP350 interacts with PPARs and LXR{alpha}, but not RXR{alpha}, in vitro. (A) Human PPAR{alpha} and firefly luciferase (Luc) as a control for nonspecific protein binding, and (B) PPAR{gamma}2, PPAR{delta}, LXR{alpha} and RXR{alpha} 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{alpha}, and 9-cis retinoic acid for RXR{alpha}. Lanes designated Load (1/10) had 10% of the L-[35S]methionine-labeled protein added to each of the respective binding assays.