Cyp2b6

Real-time quantitative RT-PCR (qRT-PCR) was performed for selected genes using Taqman chemistry and the ABI 7900 Real Time PCR System. The Globin cleared purified RNA from 5 control and irradiated animals was used for cDNA synthesis using the High-Capacity cDNA Archive Kit (Life Technologies). Gene expression assays (primer/probe sets) were purchased from Thermo Fisher for the following genes: Ifit3 (Mm00366278_m1), Ifit1 (Mm00365614_m1), Cdkn1a (Mm04205640_g1), Aen (Mm00471554_m1), Gstt1 (Mm00492506_m1), Il18r1 (Mm00515178_m1), Ccng1 (Mm00438084_m1), Alas1 (Mm001235914_m1), Acsbg1 (Mm00547366_m1), Cyp2b6 (Mn00456591_m1), Actb (Mm00607939_s1). The ΔΔCT method was used to calculate expression relative to controls, using normalization to Actb expression. Each reaction was run in triplicate with 5 control and 5 irradiated samples, and means were compared using an unpaired t-test.

Dimethyl sulfoxide (DMSO) was provided by Glentham Life Sciences (Corsham, UK). PCR-grade nano-pure water was purchased from Jena Bioscience (Munich, Germany). Vivid CYP1A2, CYP2B6, CYP2C19, CYP2D6, and CYP3A4 screening kits, EOMCC, and BOMCC were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Betanin, bupropion, furafylline, omeprazole, quinidine, rifampicin, and Krebs-Henseleit buffer were supplied by Sigma-Aldrich (St. Louis, MO, USA). Sulfaphenazole, fluvoxamine, and ketoconazole were obtained from Toronto Research Chemicals (Toronto, ON, Canada). P450-Glo CYP1A2, P450-Glo CYP2B6, and P450-Glo CYP2C9 assay kits were provided by Promega (Wisconsin, MD, USA).

RT-qPCR was used to examine CAR target gene
expression in PHH, HepaRG cells, or in mouse liver samples. Total
RNA, reverse transcription, and qPCR were performed, and mRNA expression
data was analyzed as we have described before.^{51 (link)}All RT-qPCR experiments were performed in triplicate
samples, and data are presented as fold induction to vehicle-treated
cells with the same reagents we have described before.^{48 (link),52 (link)} PCR TaqMan probes for murine genes have been listed in our previous
reports.^{52 (link)} TaqMan probes for CYP3A4, CYP2C9, and CYP2B6 human genes
as well as for reference genes B2M and GADPH genes were obtained from Thermo Fisher: CYP3A4 (Hs00604506_m1), CYP2C9 (Hs02383631_s1), CYP2B6 (Hs04183483_g1), GAPDH (Hs02758991_g1), and B2M (Hs00984230_m1).