Fig. 1

Inhibition of AR signaling limits CDC6 and induces senescence in androgen-sensitive prostate cells. A Schematic demonstrating prostate cancer development. The androgen-dependent LNCaP prostate cancer cell line represents enzalutamide-sensitive localized cancer. LNCaP-derived C4-2B prostate cancer cells are isolated from bone metastases and constitute androgen-resistant cells. PC-3 prostate cancer cells are also isolated from metastatic to bone adenocarcinoma and represent androgen-resistant cells. B Western blotting and densitometry of control and enzalutamide (ENZA)-treated LNCaP cell lysates with the indicated antibodies. Complete absence of PSA expression upon enzalutamide treatment verifies responsiveness to treatment. GAPDH was used as endogenous control. C GL13 immunocytochemical staining of control and enzalutamide-treated LNCaP cells. Lipofuscin predominantly accumulates in the cytoplasm of treated LNCaP cells. Magnification: 200x (Objective 20x), scale bars: 30 μm. See also Fig. S1A and B. D Quantification of C. E MTS proliferation assay verifying reduced proliferation of enzalutamide-treated LNCaP cells. Enzalutamide was used at a 10 μM concentration. ***P < 0.001 of Student’s t-test. Error bars indicate s.e.m. Data shown are representative of at least three biological replicates (n ≥ 3). Vectors were obtained from www.vecteezy.com