Figure 1. DEK modulates VEGF expression in breast cancer cells.

A. Schematic demonstration of screening for transcription factors that regulate VEGF-Luc reporter activity in ZR75-1 breast cancer cells. B. ZR75-1 breast cancer cells were cotransfected with the VEGF-Luc reporter and the indicated transcription factors from A. Luciferase reporter assays were performed to detect the effect of transcription factors on VEGF-luc reporter activity. All values shown are expressed as the mean ± SD obtained from two independent experiments. **P < 0.01 versus empty vector. C. ZR75-1, MCF-7 and MDA-MB-231 breast cancer cells were cotransfected with VEGF-Luc and DEK shRNA1, DEK shRNA2 or control shRNA. Luciferase reporter assays were performed to detect the effect of DEK shRNAs on VEGF-luc reporter activity. Representative immunoblot indicates the expression of DEK. GAPDH was used as a loading control. D. Real-time RT-PCR analyses of the expression of VEGF₁₂₁ and VEGF₁₆₅, two major VEGF isoforms, in ZR75-1, MCF-7, and MDA-MB-231 cells stably infected with lentivirus carrying DEK shRNAs or control shRNA. Representative immunoblot shows DEK expression. E. VEGF concentration in cell supernatants from ZR75-1, MCF-7 and MDA-MB-231 cells stably infected as in D was analyzed by ELISA assay. Data shown are mean ± SD of triplicate measurements that have been repeated 3 times with similar results (C-E). *P < 0.05, **P < 0.01 versus control shRNA.