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. Author manuscript; available in PMC: 2020 Apr 1.
Published in final edited form as: Mol Cancer Ther. 2019 Jul 15;18(10):1875–1886. doi: 10.1158/1535-7163.MCT-18-1322

Figure 4. AKR1C3 controls AR/AR-V7 protein stabilization in resistant prostate cancer.

Figure 4.

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.