Fig. 2

TNFα-mediated IFNε gene expression is mediated by the NFκB pathway. a Primary cervical cells were pre-treated with or without IKK inhibitor, BAY 11-7082 at 10μM, for 1 h followed by TNFα (1 ng/ml) stimulation. Total RNA was prepared and IFNε gene expression was measured by RT-qPCR. b Primary cervical cells were pre-treated with or without BAY 11-7082 at 10μM for 1 h before TNFα stimulation for 30 min. The Ser 536 phosphorylation of NFκB p65 was measured by Perkin Elmer AlphaLISA SureFire Ultra pNFκB p65 (Ser 536) assay kit according to the manufacture’s instruction. c A2EN cells were transfected with pGL3 or pGL3 containing IFNε promoter. The promoterless pGL3 basic vector was included as the baseline control. Cells were then treated with or without 1 ng/ml TNF for 24 h before measuring luciferase activity (RLUs). d A2EN cells were transfected with the IFNε promoter construct and then treated with or without BAY 11-7082 at 10μM for 1 h. Cells were then stimulated with or without TNFα at 1 ng/ml for 24 h before the measurement of the luciferase activity. Data are mean ± SD of three independent experiments. *p < 0.05