Androgen receptor status of the 305 and 364 cell-lines.

<p>(A–B) Cells were grown in Medium A with 10 nM testosterone (T) or 10 μM casodex (CDX). (A) Protein was harvested and subjected to SDS-PAGE and Western blotting against the androgen receptor (AR) and α-tubulin. (B) RNA was harvested and <i>PSA</i> mRNA levels were determined by qRT-PCR, normalised to the LNCaP cells. (C) <i>Top panel</i>: Cells were starved in Medium B for 24 h, before treatment with 1 nM dihydrotestosterone (DHT) and/or 10 μM CDX in Medium B for another 24 h. Following treatment, RNA was harvested and <i>PSA</i> mRNA levels were determined by qRT-PCR, normalised to the vehicle-treated LNCaP cells. <i>Bottom panel</i>: Following transfection, cells were seeded in Medium B. The next day, cells were treated with 1 nM DHT and/or 10 μM CDX in Medium B for another 24 h. Following treatment, cells were assayed for luciferase activity, made relative to the vehicle condition within each cell-line. (D) Summary of the results obtained in (A–C). (A) Blots are representative of four separate experiments. (B–C) Data presented as mean + S.E., from three separate experiments per cell-line, each performed with triplicate wells per condition.