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. 2018 Jul 10;8:10394. doi: 10.1038/s41598-018-28500-z

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Chronic treatment of BaP induces expression of CYPs in U1 cells. The U1 cells were treated with 100 nM BaP for seven days. We measured the mRNA expression (A,D) and protein expression (B,E) of CYPs (1A1, 3A4) using RTPCR and western blotting, respectively. Chronic exposure of BaP (100 nM) significantly induced the expression of CYP1A1 at the mRNA level, but not at the protein level. Therefore, we measured activity of CYP1A1 using the EROD assay (C). Chronic BaP treatment increased CYP1A1 activity by approximately 2.5-fold. However, there was no significant change in the expression of CYP3A4 at both the mRNA and protein levels. The data are displayed as mean ± SEM of at least three independent experiments (n ≥ 3). The mRNA/protein expression of the treated cells are normalized to control cells, whose expression was set at 1-fold. GAPDH was used as an endogenous control and loading control for RTPCR and western blotting, respectively. The statistical significance was calculated at *p ≤ 0.05 compared with the control group. The blots are representative of at least three independent experiments.