Cell proliferation in EAF2- and ELL2-deficient prostate cancer cells. (A) BrdU incorporation in C4-2 cells transfected with nontargeted control (siCtrl) siRNA, targeted to EAF2 (siEAF2), ELL2 (siELL2), or concurrent EAF2 and ELL2 (siDouble) knockdown. The upper panel shows BrdU-positive nuclei (red), and the lower panel shows nuclear staining with SYTOX Green (green). Original magnification, × 40. (B) Quantification of BrdU incorporation shown as mean percentage ± standard deviation of BrdU-positive cells relative to the total number of cells (at least 400 total cells were counted for each condition). (C) BrdU incorporation in 22Rv1 cells transfected with nontargeted control (siCtrl) siRNA, targeted to EAF2 (siEAF2), ELL2 (siELL2), or concurrent EAF2 and ELL2 (siDouble) knockdown as above. (D) Quantification of BrdU incorporation. Results are representative of four individual experiments. (E) Western blot analysis of EAF2 and ELL2 protein from C4-2 cell lysates and (F) 22Rv1 cell lysates following siRNA knockdown as in (A). GAPDH served as internal loading control. *P < 0.05, **P < 0.01, ***P < 0.001.