Results of the high-throughput FRET assay, selection and structure of Compound 10. (A) Summary of the high-throughput NFAT-AP1 inhibitor screen and flowchart of compound selection. (B) Structure of Compound 10 (PubChem ID: 1432799). (C) Concentration-dependent inhibition of the FRET signal by Compound 10. The results are plotted as observed FRET ratio (acceptor fluorescence/donor fluorescence) relative to the FRET ratio of wells containing the complete assay mix without Compound 10. Red symbol, control wells containing the complete assay mix (10 nM ARRE-2–Alexa-546, 10 nM Fos-OG, 10 nM Jun, and 20 nM NFAT); black symbol, control wells with NFAT omitted; blue symbols, wells with complete assay mix and the indicated concentrations of Compound 10. IC50 = ∼2 μM.