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. 2012 Oct;82(4):601–613. doi: 10.1124/mol.112.078147

Fig. 2.

Fig. 2.

Effect of electrophiles on AP-1 gene expression in mouse liver. A, BHA induces c-jun and c-fos gene expression in mouse liver. Nrf2(+/+) (n = 4) or Nrf2(−/−) (n = 3) mice on a C57BL/6 background were fed an AIN-76A diet (control) or a diet supplemented with 0.45% BHA (treated) for 7 days. qRT-PCR analysis of mRNA levels of c-jun and c-fos in mouse liver. *, p < 0.05, significantly different compared with Nrf2(+/+) control mice; **, p < 0.05, significantly different compared with BHA-treated Nrf2(+/+) mice (one-way ANOVA followed by the Tukey test.) B, C57BL/6J mice were fed either an AIN-76A diet (control, n = 4) or a diet supplemented with 0.45% BHA (fed, n = 4) for 7 days. Western blot procedures are described under Materials and Methods. The membranes were incubated with antibodies against c-Jun (1:1000) and phospho (p)-c-Jun (1:1000; Ser63) overnight at 4°C. Western blot was repeated three times, and the figure is a representative blot. C, densitometry analysis of c-Jun protein and phospho-c (pc)-Jun (Ser63) levels in mouse liver in response to BHA. *, p < 0.05, significantly different compared with control (Student's t test). D, acrolein induces c-jun and c-fos gene expression in mouse liver and Hepa-1c1c7 cells. C57BL/6J mice were treated with water (control, n = 5) or 5 mg/kg acrolein (treated, n = 5) by gavage daily for 7 days. Hepa1c1c7 cells were exposed to 20 μM acrolein for 6 h. The mRNA levels of c-jun and c-fos were assessed by qRT-PCR. *, p < 0.05, significantly different from DMSO control (Student's t test). E, time-dependent activation of c-Jun by acrolein in Hepa-1c1c7 cells. Cells were treated with 20 μM acrolein at various time intervals (0–4 h). After separation by electrophoresis, the proteins were transferred to membranes and the membranes were probed with antibodies against phosph-c-Jun (Ser63), c-Jun, JNK1/2, and phospho-JNK1/2. The Western blot experiment was repeated three times with similar results and the figure shown is a representative blot. The numbers between blots represent the densitometry analysis expressed relative to zero time expressed as fold change by the number below each blot.