Figure 5.

The Nrf2-mediated GCL gene expression by Rg3. (a) H4IIE cells were transiently transfected with siRNA directed against Nrf2 or nontargeted siRNA and subsequently treated with Rg3 (3 μg/mL) or vehicle for 12 h. The representative blots of mRNA after Rg3 treatment were assessed by RT-PCR (left). To confirm gene knockdown, the Nrf2 levels 48 h after siRNA transfection were determined in the cell lysates by western blotting. GCLC and GCLM protein expression in the total cell lysates were detected in cells stimulated with Rg3 (3 μg/mL) for 24 h (right). β-actin was used as the loading control. (b) MEFs from wild-type (Nrf2+/+) or Nrf2-deficient mice (Nrf2−/−) were treated with Rg3 (3 μg/mL) or vehicle for 12 h. The mRNA levels (left) and protein expression (right) for the genes were analyzed as described above (a). All experiments were performed using independent mRNA and lysate samples. Each value represents the average with at least three separate experiments.