No significant changes in cell cycle profile was observed in DON-treated cells

e indicated time periods and stimulated overnight with LPS or left unstimulated. After fixation in 2% PFA, cellular actin was stained with phalloidin-Alexa568 and cells were imaged on a Zeiss LSM510 meta confocal laser scanning microscope. The number of filopodia extending radially from the cell surface (expressed as filopodia per mM contour length; see M&M) was determined for control cells and cells treated for 24 hours with oligomycin, in the presence and absence of LPS (K). The average cell circumference was determined for cells in control medium or medium containing 10 LY341495 web pubmed ID:http://www.ncbi.nlm.nih.gov/pubmed/19652226 mM galactose, or 1 mM glucose and 10 mM galactose (L). (p,0.05, p,0.01, p,0.001, unpaired t-test). doi:10.1371/journal.pone.0096786.g003 Finally, we wanted to determine whether the inhibitory effect of acute glucose deprivation on phagocytosis could be directly rescued by reintroducing glucose. RAW 264.7 and Maf-DKO cells were incubated with FITC-COZ in galactose medium for 30 minutes after which 1 mM of glucose was added and cells were incubated for another 30 minutes. RAW 264.7 phagocytosis capacity was restored to 45% while Maf-DKO cells regained 75% of their phagocytosis capacity (Figure 6I). Taken together, our findings imply that glycolysis is critical for phagocytosis of COZ while OXPHOS activity is dispensable. Moreover, our results suggest that macrophages require the presence of glucose in order to successfully bind and internalize COZ particles. Discussion Increased morphodynamic activity, facilitated by rearrangement of the actin cytoskeleton, is a cellular response characteristic to LPS-stimulated macrophages. This dynamic actin remodeling is essential to macrophage function and considered to be an energy draining process. In endothelial