Figure 4.

Transcriptional activity of ARE1 and ARE2 measured in terms of luciferase reporter activity in LNCaP and C81 cells. (A) ARE1 and ARE2 either together (ARE1+ARE2), alone (ARE1 or ARE2) or mutant ARE1 (mtARE1) was cloned upstream of a minimal promoter luciferase reporter plasmid. (B) The mutant ARE1 was created by mutating the conserved “C” and G” in the ARE to “A” and “T” respectively (underlined). (C and D) The reporter plasmids were transiently transfected in LNCaP (C) or C81 (D) cells cultured in media containing CFBS and treated as indicated. The cells were also co-transfected with Renilla luciferase used as a transfection control. The luciferase activity measured 24 hrs after transfection was normalized to Renilla luciferase and expressed as relative luciferase activity. The data is expressed as mean ± SEM of 3 experiments performed in quadruplicates. The t-test significance (***P < 0.001) is measured against the activity of ARE1+ARE2 within respective treatment groups.