The IL-8 promoter contains a true RE/AP composite element.
(A) Jurkat T cells were transfected with reporters containing 4 copies of the wild-type (wt) IL-8 RE/AP element, or constructs containing mutations at the NF-κB site, AP-1 site, or both. The following day, the 105 live cells were unstimulated or stimulated with 5 ng/mL PMA and 1 μg/mL anti-CD28 (Caltag) for 6 hours as denoted in the figure. Luciferase activity was standardized to the unstimulated control (n = 1). The actual average counts for background IL-8 RE/AP luciferase activity were 1020. The results shown are the average of 3 independent transfections. Error bars reflect the SD. (B) Jurkat cells were transfected as above, except that reporter constructs containing either the wild-type IL-8 promoter or ones containing mutations at either the NF-κB site or AP-1 sites within the IL-8 RE/AP sites were used. Luciferase activity was standardized to the unstimulated control (n = 1). The actual average counts for background IL-8 promoter luciferase activity were 2256. The results shown are the average of 3 independent transfections. Error bars reflect the SD.
