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. Author manuscript; available in PMC: 2012 Oct 15.
Published in final edited form as: Cancer Res. 2011 Aug 25;71(20):6503–6513. doi: 10.1158/0008-5472.CAN-11-0532

Figure 3.

Figure 3

Basal AR activity in VCaP is AKR1C3 dependent. A, LNCaP, C4-2, or VCaP cells were immunoblotted for AKR1C3 or β-tubulin (loading control). B and C, VCaP cells stably infected with lentivirus expressing either GFP shRNA or AKR1C3 shRNA (Open Biosystems) in CSS medium were treated with 10 nM DHT or 100 nM androstenedione (Ad) for 24h, and then immunoblotted. D, VCaP or C4-2 cells in CSS medium were treated with 0, 20, or 40 μM indomethacin for 24h. E, VCaP cells in CSS medium were treated with 0, 20, or 40 μM indomethacin with or without 10nM DHT and subjected to qRT-PCR. F, mice bearing relapsed VCaP xenografts were treated with indomethacin for 2 weeks (~0.25 mg per day in drinking water) and tissue samples taken pre- and post-therapy from tumors (n=5) were analyzed by RT-PCR as indicated or (G) by immunohistochemistry for PSA, ERG, AR, and Ki67. H, testosterone and DHT levels in recurrent xenograft tumors in transplanted female scid mice (n=3) were measured pre- or post-indomethacin (Indo) treatment.