Figure 6

Mutation of both ets-binding site and CArG box abolished the activation of 136 bp length ZC3H12A promoter fragment. Luciferase activity was measured to examine the regulation of 136 bp length ZC3H12A promoter fragment. HepG2 cells were transiently transfected with the luciferase construct containing the fragment of human ZC3H12A promoter located between -76 bp and +60 bp. Black bars - wild type promoter, striped bars - promoter with the mutation in both ets-binding site and in CArG box. A, 24 h after transfection cells were stimulated with IL-1β and PMA for 3 hrs. B, Cells were co-transfected with increasing amounts of Elk-VP16 (20, 50 ng). Luciferase activity was measured 24 h after transfection. C, Influence of mutation of ets-binding site or CArG box on the activation of 136 bp by Elk-1. Black bars represent wild type promoter, dotted bars represent promoter with the mutation in ets-binding site, light bars represent promoter with the mutation in CArG box. Cells were co-transfected with increasing amounts of Elk-VP16 (10, 20, 50 ng). Luciferase activity was measured 24 h after transfection. Statistical significance was determined using the Student's test. *P < 0.05, **P < 0.001.