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. 2012 Jan 12;26(3):423–437. doi: 10.1210/me.2011-1233

Fig. 5.

Fig. 5.

FOXO1 increases NF-κB and CCL20 promoter interaction. A, NF-κB and CCL20 promoter interaction determined by ChIP assays. HepG2 cells incubated in normal culture medium were harvested for ChIP assays as described in Materials and Methods. The interaction between NF-κB subunits (p65, RelB, C-rel, p52, and p50) and CCL20 promoter was determined. B, Interaction between NF-κB proteins (p65, p50 or p65/p50) and CCL20 promoter. HepG2 cells treated with TNF-α (10 ng/ml) for 24 h were harvested as described in Materials and Methods and immunoprecipitated with anti-p65, anti-p50, or both antibodies. C, FOXO1 stimulation of NF-κB (p65 and p50) and CCL20 promoter interaction determined by ChIP assays using HepG2 cells transfected with FOXO1-AAA or pcDNA3.1 plasmid for 24 h, followed by 24-h treatment of TNF-α (10 ng/ml). D, FOXO1 silencing inhibits NF-κB (p65 and p50) and CCL20 promoter interaction as determined by ChIP assays using HepG2 cells transfected with FOXO1 siRNA (siFOXO1) or a scrambled siRNA (siNC) for 24 h, followed by 24 h of TNF-α treatment (10 ng/ml). Each of the experiments was repeated two to four times, and representative results were displayed.