Mold Alkaloid Cytochalasin D Modifies the Morphology and Secretion of fMLP-, LPS-, or PMA-Stimulated Neutrophils upon Adhesion to Fibronectin

Figure 4

SDS-PAGE separation of proteins secreted by neutrophils upon adhesion to fibronectin in the presence of fMLP and cytochalasin D. Human neutrophils were attached to fibronectin-coated substrata for 20 min in the presence of 1 μM fMLP taken separately (a) or in combination with 10 μM cytochalasin D (b). Samples of extracellular medium were collected and proteins were extracted and, after concentration, were subjected to 15% SDS-PAGE. The gels were stained with Coomassie Brilliant Blue. The lines mark the places from which pieces of gel were cut. In these pieces, after trypsin hydrolysis directly in the gel, the designated proteins were detected by mass spectrometry. Pictures represent typical protein profiles, observed in three independent experiments for each EM example.