Figure 1
From: Characterization of a new selective glucocorticoid receptor modulator with anorexigenic activity
Selective inhibition of Agrp-GRE by a natural steroid compound. (A) Schematic representation of the Agrp promoter containing binding sites for Bsx and GR. Agrp-GRE, MMTV-GRE and consensus GRE are highlighted. Asterisks indicate the sequences in the first half site (GGA) and the region flanking the second half site (GTTTC) of Agrp-GRE that are critical for the unique properties of Agrp-GRE (i.e., synergy with Bsx in the induction of orexigenic genes). (B) Structure of the Zj7 compound. (C–I) Luciferase reporter assays with AgRP-1 kb:LUC, (Agrp-GRE)3:LUC, PER1:LUC, MMTV:LUC and MT1-LUC in HEK293T cells. AgRP-1 kb:LUC was synergistically activated by Bsx and GR treated with the synthetic GC, dexamethasone (Dex, 10−7 M). Treatment of naturally derived steroidal compound Zj7 suppressed AgRP-1 kb:LUC, (Agrp-GRE)3:LUC and PER1:LUC, but did not affect MMTV:LUC and MT1-LUC. RU486 inhibited both AgRP-1 kb:LUC and MMTV:LUC activated by GR and Dex. Transfections were repeated at least three times independently. Data are presented as the mean of triplicate values and error bars represent the standard deviation. Statistical differences were determined by Student’s t-test; *p < 0.05, and **p < 0.01 and ‘ns’ indicates not significant. (J) Zj7 negatively targets non-classical Agrp-GRE but not classical GRE.
