Figure 5
From: Regulation of the arachidonic acid mobilization in macrophages by combustion-derived particles
MAF02-induced mobilization of AA depends on ERK1/2 and MAPKs are activated in RAW264.7 macrophages as well as in human MDM. (A) RAW264.7 cells labelled with [14C]arachidonic acid were pre-incubated for 30 min with the indicated kinase inhibitors and subsequently exposed to 50 μg/ml MAF02 particles (12.6 μg/cm2) for 2.5 h. After lipid extraction, free AA was separated by TLC, visualized by autoradiography, and the optical density of the bands were analyzed by OptiQuant® and Odyssey® software. [14C]arachidonic acid mobilization data are expressed as a percentage of the values for control cells (100% values). Results are the mean ± s.e.m. of three independent experiments (***p < 0.001 compared to 50 μg/ml MAF02-treated cells). RAW264.7 macrophages (B) and human MDM (C) were treated with 50 μg/ml MAF02 particles (15.6 μg/m2) for 1, 2.5 and 5 hours and whole cell lysates were analyzed for the target proteins by Western blotting. The respective non-phosphorylated proteins are used as loading control.