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. 2014 Mar 15;28(6):548–560. doi: 10.1101/gad.237081.113

Figure 3.

Figure 3.

Circadian rhythm of NRF2 binding to ARE drives rhythmic oscillations of genes involved in glutathione synthesis and utilization. (A) Temporal mRNA levels of NRF2 targets in wild-type (WT) mouse lungs (DD). Data (mean ± SEM) were normalized to Gapdh, and the lowest expression was set as 1. (*) P < 0.05, one-way ANOVA. (B) Rhythmic NRF2 occupancy on AREs of antioxidant gene promoters in wild-type mouse lungs by NRF2-specific ChIP. The position of each ARE in relation to the TSS is shown. Mean ± SEM. (*) P < 0.05; (**) P < 0.01, t-test. (White bars) Light phase; (black bars) dark phase. (C) Temporal mRNA profiles of NRF2 target genes and clock gene Dbp in MEFs from wild-type and Nrf2 knockout (KO) mice following serum shock. Harvest of cells started at 28 h after serum shock. Data (mean ± SEM) were normalized to Gapdh. (*) P < 0.05 for all six genes in wild type; not significant for Nrf2 knockout MEFs (except Hmox1 and Dbp; [*] P < 0.05), one-way ANOVA for the effect of time. (D) Temporal levels of reduced glutathione (GSH) in MEFs from wild-type and Nrf2 knockout mice following serum shock. Data (mean ± SEM) were normalized to cellular counts and quantified using standard curve for reduced GSH. (**) P < 0.01; (ns) not significant, t-test.