Fig. 2.

Antiandrogen activity of the D4 phenothiazine derivative. The CAT reporter gene assays were conducted as described in Materials and Methods. (A) Inhibition of DHT (1 nM)-stimulated CAT activity, in CV-1 cells transfected with the wild-type AR, by D4 (○) or Casodex (●). CAT activity data are normalized against β-gal activity. (B) D4-stimulated CAT activity in CV-1 cells transfected with the T877A mutant AR. (C) Effect of increasing concentrations of D4 on β-gal activity in wild-type AR-transfected CV-1 cells. (D) PSA assay in LNCaP cells. (−), untreated cells (negative control); (+), 50 nM DHT alone (positive control). The ligand was added to the cells at the indicated concentrations together with 50 nM DHT. The films were digitized, and PSA expression data were normalized by α-tubulin content. (Inset) Endogenous AR expression levels in LNCaP cells. The data are mean ± SEM of three independent experiments. ∗, P <0.05. a.u., arbitrary units.