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. 2016 Jun 14;6:27876. doi: 10.1038/srep27876

Figure 3. Effects of cecropin B on the transcriptional activity of CYP3A29 and PXR.

Figure 3

(A) Cells were treated with 500 ng/ml of cecropin B for 12 hours in the absence or presence of 5 μM of DRB. Total RNA was prepared and CYP3A29 expression levels were analyzed by qRT-PCR. B: The luciferase reporter gene of CYP3A29 for cecropin B-regulated gene expression analysis. (B), left) HepLi cells were transiently cotransfected with CYP3A29 promoter reporter plasmids. (B), right) After 24 hours, transfected cells were treated with cecropin B. Luciferase activity was assayed 12 hours after treatments; ** and * P < 0.01 and P < 0.05, respectively. (C) Prediction of the PXR/RXR-α binding site in the CYP3A29 promoter region using JASPER software. D: HepLi cells were transiently cotransfected with site-specific mutant CYP3A29 promoter reporter plasmids. After 24 hours, transfected cells were treated with cecropin (B). Luciferase activity was assayed at 12 hours after the treatments. E: ChIP assay was performed using chromatin from Hep Li cells and anti-RNA polymerase II (positive), normal mouse IgG(negative), anti-PXR(PXR) were used as the immunoprecipitating antibody to immune select for the DNA for protein of interest that is specifically complexed with it. Purified DNA and total DNA (input) were then analyzed by PCR using primers specific for the C region.