Compound JJ-450 inhibits ARF876L-mediated cell proliferation. Representative images show BrdU staining in C4–2 GFP-ARWT (A) or C4–2 GFP-ARF876L (B) sublines. C4–2 GFP-ARWT or C4–2 GFP-ARF876L stable sublines were treated with DMSO, 5 μM Enz, 10 μM Enz, 5 μM JJ-450 or 10 μM JJ-450 in complete medium for 48 h prior to BrdU assay as described in materials and methods. Upper panel shows nuclear staining with SYTOX Green (green), middle panel shows BrdU-positive nuclei (red), lower panel shows merged images of SYTOX and BrdU staining. (C) and (D) show quantification of BrdU-positive cells percentages of C4–2 GFP-ARWT and C4–2 GFP-ARF876L subline, respectively. Data represent mean ± SD, relative to the control. *P<0.05; **P<0.01; ***P < 0.001. Enz, enzalutamide.