Fig. 4
From: A structural mechanism for directing corepressor-selective inverse agonism of PPARγ
Mutagenesis validates the R288-pyridyl interaction for conferring corepressor-selective inverse agonism. a HEK293T cells transfected with full-length PPARγ expression plasmid along with a 3 × -PPRE-luciferase reporter plasmid and treated with the indicated ligands (5 µM). Individual points (n = 6) normalized to DMSO control (mean) are plotted on top of a box-and-whiskers plot; the box represents 25th, median, and 75th percentile of the data, and the whiskers plot the entire range of values. b, c Affinities determined from a fluorescence polarization assay of wild-type and mutant PPARγ LBDs preincubated with a covalent ligand (GW9662 or T0070907) or vehicle (DMSO) binding to FITC-labeled b TRAP220 or c NCoR1. Data plotted as the Kd value and error from fitting data of two experimental replicates using a one site binding equation. d Legend to the web of efficacy radar chart diagrams. Corepressor-selective inverse agonism is associated with data points populating the periphery of the plots. e Radar web of efficacy plots displaying assay data for wild-type PPARγ and mutant variants. Data normalized within the range of values for each assay (a–c). Data are representative of at least 3 independent experiments
