Figure 1: The effect of I281T mutation on CAR activity. (A) Activation of PBREM-driven reporter by empty vector (white columns), wild-type CAR (black columns) or I281T mutant (grey columns) full-length receptors. (B) Activation of GAL4-driven reporter by empty vector (white), wild-type CAR (black) or I281T mutant (grey) LBD constructs. (C) Activation of PBREM-driven reporter by empty vector (white), wild-type CAR (black) or I281T mutant (grey) full-length receptors in the absence or presence of co-transfected coactivator PGC1α. (D) Activation of PBREM-driven reporter wild-type CAR (black) or I281T mutant (grey) full-length receptors after a 24-hour exposure to either vehicle DMSO, agonist CITCO (1 μM) or inverse agonist S07662 (10 μM). The data shown in A - D are means ± SEM from three independent experiments, each with three biological replicates and the results were normalized to the activity of wild-type CAR (set at 100). A and B shows each independent experiment (Expt #1 - #3) while C and D show combined results. * denotes a statistically significant difference (p < 0.017) to empty vector (A – C) or wild-type CAR exposed to DMSO (D). # denotes a statistically significant difference (p < 0.017) to wild-type CAR (A – D).