Fig. 1

Effects of BRD4 knockdown on urothelial carcinoma cell lines. a Western blot analysis of BRD4 expression in 10 UCCs compared to the benign urothelial control cell lines HBLAK, NHUC, and NHUC-TERT. α-tubulin served as a loading control. b Western blot analysis of BRD4 and c-MYC expression after siRNA-mediated knockdown of BRD4 in VM-Cub1 and UM-UC-3 cells. Cells transfected with BRD4 expression plasmid served as a positive control, with 3 μg instead of 20 μg protein loaded. c Relative viability of VM-Cub1 and UM-UC-3 cells after BRD4 knockdown for 48, 72, and 120 h compared to treatment with control siRNA. Relative viability is displayed on the ordinate in percent of the control cells treated with non-targeting siRNA. Differences between control and targeting siRNA were analyzed using Student’s t test (***p ≤ 0.001). d Clonogenicity assays of VM-Cub1 and UM-UC-3 cells after BRD4 knockdown for 120 h and results of quantification (**p ≤ 0.01). e Flow cytometric cell cycle analysis of VM-Cub-1 and UM-UC-3 cells after BRD4 knockdown for 48, 72, or 120 h. Percentages of cells in the respective cell cycle phase are given