Figure 7

GNMT interacts with the promoters of the genes encoding Nrf2, CAR and PXR, nuclear transcription factors. (a) The chromatin was immunoprecipitated with mouse anti-GNMT antibody (14-1) or mouse IgG (negative control). ChIP-enriched DNA was amplified with the primers for XREL1, XREL2 and XREL3 regions on the promoter of Nrf2, PXR, and CAR genes. Schematic represents the upstream promoter binding regions of XREL1, XREL2 and XREL3 in the Nrf2 (XREL1: −712 to −708; XREL2: +755 to +759; XREL3: +870 to +874), CAR (XREL1: +102 to +106; XREL2: −987 to −983) and PXR (XREL1: −1017 to −1013) genes. Horizontal arrows indicate the location of primers used for qPCR in site-specific ChIP assays. Note that the figure is not drawn to scale. (b–e) ChIP-qPCR analyses were performed with the liver lysates from AAI- or corn oil (C)-treated WT (b, female, F; c, male, M) and hGNMT Tg mice (d, female, F; e, male, M), as described in Figs 1 and 2. (f) ChIP-qPCR analyses were performed with the liver lysates from 3-week AAI- and corn oil (C)-treated WT female mice, as described in Fig. 4a (AAI 2 or AAI 5: 2 or 5 mg/kg BW/day AAI administration). The quantity of DNA in the precipitation with GNMT antibody was normalized to the IgG control. Values are presented as the mean ± SEM, n = 4. p-values were analyzed using the one-way ANOVA or Student’s t-test. *p < 0.05, **p < 0.001, ***p ≤ 0.0001.