Figure 2: Allosteric inhibition.
From: Identification of an allosteric binding site for RORγt inhibition
(a) TR-FRET assay showing the effect of the agonist cholesterol, the inverse agonist T0901317 and indazoles MRL-871, MRL-058 and MRL-003 on cofactor recruitment to the RORγt LBD in a dose-dependent manner. MRL-871, MRL-058, MRL-003 and T0901317 function as inhibitors, while cholesterol promotes cofactor binding to RORγt. Error bars are defined as s.d. (n=3). (b) IL17a mRNA expression in EL4 cells treated with MRL-871, MRL-058, MRL-003 (10 μM, 24 h) or DMSO. The level of IL17a mRNA expression was normalized to that of GAPDH expression. All data are expressed as the mean±s.e.m. (n=6). Statistical analysis was performed using an one-way analysis of variance comparing against the DMSO control following Dunnett post hoc test. Error bars denote s.e.m. **P<0.01. (c) Inhibition of cofactor binding by the allosteric modulator MRL-871 is independent of cholesterol concentration (IC50 4.7±1.4 nM; 2.5±0.9 nM; 1.8±1.5 nM; 2.3±1.8 nM at 0, 1, 10 and 25 μM cholesterol, respectively). Error bars are defined as s.d. (n=3). (d) Inhibition of cofactor binding by ligand T0901317 depends on cholesterol concentration (IC50 11±3.4 nM; 18±4.9 nM; 195±59 nM; 548±152 nM at 0, 1, 10 and 25 μM cholesterol, respectively). Error bars are defined as s.d. (n=3).
