A. C4–2B neo and C4–2B AKR1C3 cells were treated with 50 μg cycloheximide. Whole cell lysates were collected at 0, 2, 4 and 8 hours after treatment and subjected to western blot. Half-lives of AR-V7 and AR-FL were calculated. B. CWR22Rv1 cells were infected with lenti-vector or lenti-shAKR1C3 particles for 48 hours and then treated with 50 μg cycloheximide. Whole cell lysates were collected at 0, 2, 4 and 8 hours after treatment and subjected to western blot. Half-lives of AR-V7 and AR-FL were calculated. C. C4–2B MDVR cells were infected with lenti-vector or lenti-shAKR1C3 particles for 48 hours and then treated with 50 μg cycloheximide. Whole cell lysates were collected at 0, 2, 4 and 8 hours after treatment and subjected to western blot. Half-lives of AR-V7 and AR-FL were calculated. D. C4–2B MDVR cells were treated with different doses of indomethacin for 3 days. Whole cell lysates were collected and subjected to western blot. E. C4–2B MDVR cells were treated with indomethacin for 3 days and then treated with 5 μg/mL MG132 for additional 6 hours. Whole cell lysates were collected and subjected to western blot. F. C4–2B MDVR cells were treated with indomethacin for 3 days, then treated with or without 5μM MG132 for additional 6 hours. Whole cell lysates were collected and immunopreciptated with AR-V7 antibody and blotted with anti-Ub and AR-V7 antibodies. Indocin: Indomethacin.