FIGURE 1.

Coculture with inflammatory macrophages induces Nrf2 activity in NCM460 cells. Macrophages deriving from monocytes treated with GM-CSF (M-g) or M-CSF (M-m) were used for coculture with NCM460 cells. A, nuclear (n.e.) and cytoplasmic (c.e.) extracts from NCM460 cells cultured for 2 days either in the absence (mono) or presence of these macrophages were submitted to Nrf2 immunoblotting, using Hsp90 and tubulin as loading control. A representative blot (left panel) and the normalized Nrf2 band intensities (right panel) as determined by band densitometry of blots from four independent experiments are shown (means ± S.D.). *, p < 0.05. B, NCM460 cells were transfected with the Nrf2-responsive pARE or the control (pGL3) firefly luciferase reporter vector together with the ptkRL reference vector expressing Renilla luciferase. Afterward, transfectants were mono- or cocultured for 2 days, and NCM460 cell lysates were analyzed for firefly and Renilla luciferase expression. Luminescence induced by firefly luciferase was normalized to the luminescence induced by Renilla luciferase; the data represent the means ± S.D. of four independent experiments performed in duplicate. RLU, relative light units.