Figure 2

Effect of anisomycin and U0126 on AA release, ERK inhibition, and cPLA ₂ Ser⁵⁰⁵ phosphorylation EGFP-cPLA₂-transfected cells were incubated with 25 ng/ml anisomycin for 30 min, 10 μM U0126 for 15 min, or with both drugs prior to stimulation with 100 μM ATP, 1 μM IONO, or 10 μM IONO. Protein extracts were subjected to immunoblotting using (A) anti-phospho-p38, (B) anti-phospho-ERK, and (C) anti-cPLA₂ antibodies. (D) AA release in response to 100 μM ATP, 1 μM IONO, or 10 μM IONO in the presence and absence of the inhibitors. Anisomycin did not stimulate AA release in unstimulated cells. The results show average ± SEM for 3 to 5 independent experiments.