Skip to main content
. 2015 Mar 6;36(4):459–468. doi: 10.1093/carcin/bgv017

Figure 2.

Figure 2.

Fra-1 is transcriptionally regulated by STAT3 in response to IL-6 stimulation. (A) Western blots with the indicated antibodies from HT-29 cells pretreated for 1h with LY294002, U0126, or Stattic (specific inhibitors of PI3K, MEK and STAT3, respectively) and exposed to IL-6 for 12h. (B) Western blots of Fra-1 and STAT3 from HT-29 cells transfected with control or STAT3 siRNAs and incubated with IL-6 for 12h (GAPDH was used as a loading control). (C) Schematic representation of Fra-1 promoter with seven potential SIEs and the primer pair used in ChIP-PCR assays. The reporter construct Fra-1-Luc and its truncated and mutated derivatives are also shown. (D) Transcription activity in response to IL-6 treatmemt for 6h measured by luciferase assay in 293T cells with a series of deletion mutants of Fra-1-luc (internal control, pRL-TK). *P < 0.05. (E) Relative luciferase activity 6h after IL-6 incubation in 293T cells transfected with the wild-type or SIE mutated Fra-1 promoter reporter construct. *P < 0.05. (F) Chromatin prepared from HT-29 cells stimulated with IL-6 for 1h was immunoprecipitated with the indicated antibodies; PCR was performed on immunoprecipitated DNAs or soluble chromatin using specific primer pair for the Fra-1 promoter.