FIG. 5.

JDP-2 increases ligand-dependent PR-mediated transactivation. (A) Cos-1 cells were transfected with phPR-B (1.5 ng) and the progesterone-responsive reporter PRE₂-TATA-Luc (200 ng) in the presence or absence of increasing amounts of pCR3.1-SRC-1 or pCDNA3-hisJDP-2 (25, 50, 100, and 200 ng). Cells were treated with the vehicle or 10 nM R5020 for 24 h prior to harvest. Relative luciferase activity was calculated by setting normalized luciferase activity of the reporter alone to 1.0 and all other treatment group values as severalfold relative to 1.0. Values are averages ± SEM of three independent experiments. (B) Cos-1 cells were cotransfected as described for panel A above with PR-B and JDP-2, except for the use of a different progesterone-responsive reporter gene, MMTV-Luc. (C) HEC-1-B cells were transfected with phPR-B (2.5 ng) and PRE₂-TATA-luc (200 ng) in the presence or absence of pCR3.1-SRC-1 (50 ng) or pCR3.1-JDP-2 (34 ng) and were hormone treated as described above. Severalfold hormone induction was calculated as the ratio of the relative luciferase activity of hormone-treated samples divided by the relative luciferase activity of corresponding vehicle-treated samples. Values are averages ± SEM of three independent experiments. (D) Cos-1 cells were transfected with indicated nuclear receptors [phPR-B (1 ng), pRSV-GR (2 ng), SVMT-ER (0.1 ng), TRβ (0.5 ng), or RSV-VDR (1.5 ng)], together with 200 ng of their cognate hormone-responsive luciferase reporter genes [PRE₂-TATA-luc (PR and GR), ERE₃-TATA-luc (ER), TRE-luc (TR), or VDRE(1,2)-ΔMTV-luc (VDR)] in the presence or absence of pCR3.1-JDP-2 (110 ng). At 24 h prior to harvest, cells were treated with the appropriate ligands: 10 nM R5020 (PR), 100 nM dexamethasone (GR), 10 nM estradiol (ER), 100 nM T₃ (TR), or 10 nM 1,25-vitamin D₃. Severalfold hormone induction was calculated as described for panel C, and values are averages ± SEM of three independent experiments.