CCL20 promoter activity is tightly regulated by NF-κB signaling.

<p>(A) DNA sequence of human CCL20 promoter. (B) Increased effect of CXCR2 and blockage of IκB on luciferase activity of CCL20 promoter in parental SKOV-3 cells. Results were normalized to the protein level and expressed as a fold increase compared to empty vector (pA) controls. (C) Confirmation of CCL20 promoter activity and its mutants activity in response to TNF (10 ng/ml) in parental SKOV-3 cells. Site-directed mutants were generated from the CCL20-376/20LUC using primers with mutant κB-like sites (termed m1, m2 and m3 indicates mutation site): -219/-210 mutant κB-like site (CCL20LUCm1), -92/-83 mutant κB-like site (CCL20LUCm2) and -33/-24 mutant κB-like site (CCL20LUCm3). After transfection with CCL20 luciferase vectors overnight, a luciferase assay was performed at post-treatment of TNF (10 ng/ml) for 6 h. Results were normalized to the protein level and expressed as a fold increase compared to non-treated control (C). Cross circles indicate κB site mutants. pA = empty vector transfection; pCXCR2 = CXCR2 vector transfection. *,^# indicate significant (p≤0.05) increase and decrease, respectively, when a Student’s-<i>t</i> test was analyzed.