Skip to main content
. 2013 Sep 6;5(6):391–403. doi: 10.1093/jmcb/mjt035

Figure 2.

Figure 2

E2F1 directly interacts with human VEGFR-3 and VEGF-C promoters. (A) Relative luciferase activities measured 30 h after cotransfection of VEGFR-3−849/+55 or VEGF-C−49/+419 promoter construct with increasing amounts (0.5, 1, 2, and 3 µg) of E2F1 expression plasmid or DNA-binding-deficient E132 mutant (3 µg). Western blots confirm E2F1 and E132 protein expression after transfection (right). Actin was used for equal loading. (B) Schemes of VEGFR-3 and VEGF-C promoter deletion constructs with relative locations of predicted transcription factor binding sites. Luciferase activity was measured in response to E2F1 overexpression. (C) ChIP in SK-Mel-29.ER-E2F1 cells cultured 24 h in the presence or absence of 4OHT using anti-E2F1 or IgG. Input represents 10% of sheared chromatin prior to immunoprecipitation (left). E2F1-dependent VEGFR-3 and VEGF-C promoter reporter activities were detected in the absence or presence of 4OHT (right). (D) Cotransfection of VEGFR-3−849/+55 or VEGF-C−49/+419 promoter reporters and different E2F family members. E2F1, E2F2, and E2F3a expression was verified by western blotting. Promoter activity is normalized to per mg protein and relative to empty vector.