Fig. 5

AP1 and CRE response elements do not mediate GR transrepression. a, Sequence analysis of a short fragment upstream of exon IV, and included in SP4 sequence that contains two potential AP1 binding sites, TGTATCA and TGATTCT, as well as two potential cAMP Response-Elements (CRE): CRE-1 (for CREB1, TCACGTCA) and CRE-2 (for CREB3l2, TCTCGTGG) according to the Jaspar database; +1 is the transcription start site of exon IV. b, Luciferase activity of WT and AP1 mutants. mAP1-1: construct with AP1-1 deletion, mAP1-2, construct with deletion of AP1-2 sequence, dmAP1, deletion of both sites. Deleted based are shown in white lettering in Fig. 5a. Basal transcriptional activity driven by the WT sequence was arbitrarily set at 1, Means ± SEM; *** P < 0.005; Mann Whitney U-tests, n = 8. c, Luciferase activities of promoter mutants with AP1 deletions were still repressed by DEX. * P < 0.05 DEX (red) vs vehicle (blue). Mean ± SEM; ** P < 0.01, DEX vs Veh of the same construct. Mann Whitney U-tests, n = 8; all Veh conditions were set at 1. d, Luciferase activity of WT and CRE mutants. mCRE-1: construct with CRE1-1 deletion, mCRE-2, construct with deletion of CRE-2 sequence, dmCRE, deletion of both sites. Deleted based are shown in white lettering in Fig. 5a. Basal transcriptional activity driven by the WT sequence was arbitrarily set at 1, *** P < 0.005; Mann Whitney U-tests, n = 8. e, Luciferase activities of promoter mutants with CRE deletions were still repressed by DEX. mCRE-1: construct with CRE-1 deletion, mCRE-2, construct with deletion of CRE1-2 sequence, dmCRE, deletion of both sites. * P < 0.05 DEX (red) vs vehicle (blue). Means ± SEM; ** P < 0.01, DEX vs Veh for the same construct; *** P < 0.005, DEX vs Veh for the same construct, Mann Whitney U-tests, n = 8; all Veh conditions were set at 1