It is well documented that KH domains bind only to regions of single stranded RNA

ent-opsonized Zymosan by LPS-stimulated Macrophages is Fueled by Glycolysis and Requires the Presence of Extracellular Glucose We finally examined whether metabolic conditions (with differential nutrient supply or inhibitor conditions as above) also affect phagocytosis, the main actin-dependent function of macrophages [27]. Formation of the phagocytic cup is driven by local polymerization of actin filaments, associated with membrane alterations and regionally confined topological alterations at the cell surface. Unlike cell spreading, particle uptake via phagocytosis, therefore, is localized and determined by mechanistic events that occur within the small area of contact between cell and particle. Acute inhibition (0 h pre-incubation) of OXPHOS initially led to a reduction in internalization efficiency as well as the overall phagocytic index of RAW 264.7 cells (Figure 6A, B), however, after three hours, phagocytosis efficiency was restored and with longer oligomycin treatment (24 h) phagocytic index and the particle internalization capacity was even higher than in control cells. In the presence of ample glucose (25 mM), competitive inhibition of glycolysis by 2-DG only inhibited phagocytosis significantly after pre-incubation (Figure 6C). 2-DG did not have an acute effect, but after 1 hour of 2-DG pre-incubation phagocytosis was markedly down (p = 0.055). This effect became PLOS ONE | www.plosone.org 7 even more pronounced with prolonged (3 h) pre-treatment, a period wherein cell viability was still not affected (Figure 1D). The efficiency of particle internalization was, however, not compromised (Figure 6D) by addition of 2-DG. Phagocytosis in the presence of oligomycin PF-562271 custom synthesis PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19651432 and 2-DG was also assessed in a second macrophage