A). To assess AR activity on cyclin D1 transcript-a and transcript-b levels, LNCaP cells were cultured in either in the absence of steroid hormones (charcoal dextran treated serum, CDT) and stimulated 24 hrs with (or not) with 1 nM DHT, as indicated, or in complete serum (5% FBS) supplemented where indicated with 1 uM Casodex. Real time PCR for relative KLK3/PSA (upper panel), transcript-a (middle panel), transcript-b (lower panel) are shown. Representative images are shown in Supplemental Figure S3. B). To evaluate the contribution of mTOR-mediated post-transcriptional regulation of cyclin D1b, LAPC4 cells were treated with 10 uM RAD001 for 4 hrs. Cyclin D1b levels were determined by immunoblot analysis using the cyclin D1b-specific antibody with lamin (control). Representative immunoblot is shown.