ROS, Nrf2 and GSH levels, and γ-GCS activity in iB16 and iB16-shGCR cells isolated from metastatic foci.

Melanoma cells were isolated from the liver 7 days after inoculation, cultured, and transfected with anti-Nrf2-siRNA. H2O2 and O2−generation, γ-GCS activity, and GSH levels were measured 48 h after seeding. Nrf2 levels (Western blotting) were measured 24 h after seeding. AU, arbitrary units. Data are mean values ± S.D. (n = 6–7 in all cases). *pversus iB16 controls. Results obtained in cells transfected with control Nrf2 sense or scrambled oligonucleotides were not significantly different from those obtained in cells cultured in the absence of anti-Nrf2-siRNA (not shown).