Figure 6. 15dPGJ₂-promoted ERK1/2 phosphorylation is dependent on oxidative stress in human neutrophils.

Neutrophils were preincubated at 37°C with or without 10 µM DEM, 100 µM Vit E or 1 mM GSH for 1 h (B). Then the cells were treated or not with 10 µM 15dPGJ₂ for the indicated times (A) or for 30 min (B, C and E), and then with 100 nM PMA for 7 min (A and B), or else with 10 µM PGD₂, 15 µM PGA₁ or 25 µM PGA₂ for 30 min (C), with 10 µM PGD₂ for 20 h (D), or with 1 µM T0901317 for 30 min (E). Phosphorylated ERK1/2 (pERK1/2) levels were analyzed on cell lysates by Western blotting and are given in arbitrary units. Immunoblots of total ERK1/2 (A-D) or GAPDH (E) are also shown for the sake of loading controls. Each blot is representative of a set of three separate experiments yielding similar results.