Figure 5. Secondary screening of selected hits from diverse Enamine compound screen.

A. Counter screen using mammalian two hybrid assay. HEK-293T cell were transfected with Gal4-REV-ERBα and VP16-NCoR as described in Figure 1. B and C. Secondary screen assay using a mammalian single hybrid assay (without any VP16 construct). B. Cells were transfected with Gal4-hPXR. C. Cells were transfected with Gal4-hCAR. Transfected cells were treated for 24 h with DMSO (0.1% v/v) or 10 µM of selected compounds and luciferase assay were performed as described above. Results are expressed as mean ± SEM and statistical significance were assessed using GraphPad prism software by a 1-Way ANOVA followed by a Tuckey post-hoc test, *P < 0.05, **P < 0.01, ***P < 0.001.