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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.

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