Figure 3

MOZ-TIF2 inhibits the transcriptional activation of ER and AR. A. MOZ-TIF2 (MT2) and ER activity. pVit-TKSL, an estrogen response element-driven luciferase reporter plasmid was co-transfected into CV-1 cells with an estrogen receptor expression vector and pCDNA3.1-MOZ, pCDNA3.1-MOZ-TIF2 (MT2), pCDNA3.1-TIF2 or vector alone. B and C. MOZ-TIF2 and AR activity. Two androgen response element-driven luciferase reporter systems were employed. B shows the effect of a plasmid that contains a full length PSA promoter (PL) with multiple androgen response element sites and C shows the effect of a plasmid containing a minimum PSA promoter (PC) with only two androgen response elements. Both ARE-containing reporter plasmids were co-transfected with an androgen receptor expression vector and either pCDNA3.1-MOZ, pCDNA3.1-MOZTIF2 (MT2), pCDNA3.1-TIF2 or vector alone. 5α-dihydrotestosterone (DHT) was added to a final concentration of 50 nM, the cells collected and luciferase activity measured as described above. Double or single stars represent a significant difference at P < 0.01 or P < 0.05 level respectively by the two tail Student T- test compared to the transfection with MOZ-TIF2 in the ligand added condition. Open bars, 50 nM estradiol (A) or DHT (B and C). Dark bars, no added estradiol (A) or DHT (B and C). The numbers on top of the open bars, i.e. added ligand, are the ratios of light units in presence of the ligand to light units in the absence of ligand. The percentage inhibition noted in the text was calculated from the percent of light units resulting from induction by estrogen in MOZ-TIF2 expressing cells compared to pcDNA-transfected control cells subtracted from 100%.