Fig. 3. ADT downregulates REST in vitro and in vivo.

A, B Western blotting of REST in LNCaP and C4-2 cells treated with ADT drug MDV3100 (10 µM, 72 h), androgen DHT (25 nM, 24 h) or both. C Western blotting of whole cell lysates from C4-2 cells growing in media with regular FBS or CSS (charcoal stripped serum) for 7 days. Culturing in CSS media that is deprived of hormones is another common ADT approach in vitro. D Morphology of C4-2 cells growing in media with regular FBS or CSS. E RT-PCR and DNA gel electrophoresis of REST and loading control RPS18 in LNCaP xenograft tumors growing in uncastrated or castrated NOD/SCID male mice.