Differential Expression of the Activator Protein 1 Transcription Factor Regulates Interleukin-1ß Induction of Interleukin 6 in the Developing Enterocyte (original) (raw)
Fig 7
The IL-6 promoter AP-1 and NFĸB sites are equipotent in IL-1β induction of the IL-6 reporter in the immature H4 enterocyte.
**(**A) Linear map of the IL-6 promoter (IL-6 651 Luc) reporter showing the AP-1 site (-283), the NFkB site at (-72) and the C/EBP site at -154 from the transcription start site. (B) Effect of mutation of the AP-1 and NFκB binding sites on IL-1β induction of the IL-6 promoter. H4 cells were transfected with the IL-6 promoter reporter plasmids, IL-6-651 wild type, (WT) and (IL-6 Mt AP-1) mutated in the AP-1 site and (IL-6Mt NFκB) mutated in the NFκB site (C) Effect of IL-1β stimulation on NFκB and (D) AP-1 Luciferase reporter plasmids in immature H4 cells. Reporter plasmids were transfected into H4 cells together with a Renilla luciferase plasmid for estimation of transfection efficiency. After overnight culture, transfectants (n = 3) were either untreated (open bars) or treated with IL-1b (black bars) for 18h and were harvested and assayed for firefly and renilla luciferase. Data is normalized to renilla luciferase and expressed as Means +/-SE (n = 3), control un induced compared to IL-1 induced, ***p<0.001, *p<0.05. A representative experiment of three separate experiments is presented with consistent results.