Briefly, cells were lysed in ice-cold lysis buffer and 150 g of total protein was used for the assay

ivation of PAK1 and then Rac1 (another small GTPase involved in actin cytoskeletal remodeling) within 15 minutes after glucose exposure [79]. Finally, Uenishi et al. [80] have recently shown that glucose activates N-WASP via Cdc42 and induces its translocation to the cell membrane of insulin-secreting clonal pancreatic b-cells (MIN6-K8 b-cells). Moreover, glucose stimulation caused LIMK1-mediated phosphorylation and deactivation of cofilin via Cdc42 and PAK1. The timing of these effects may explain why acute removal of glucose from the culture medium had a markedly inhibitory effect on phagocytosis and also why reintroduction of glucose instantaneously restored phagocytosis capacity in our study. Interestingly, LPS signaling to the cytoskeleton also involves the Cdc42-PAK1-Rac1-LIMK1-pathway [81]. LPS stimulation additionally increases glucose uptake and metabolism via PI3K/ Akt which are signaling molecules upstream of Cdc42 and Rac1 [82�85]. Therefore, by incorporating glucose as signaling molecule, the metabolic changes that are induced by LPS may serve to reinforce the functional changes that macrophage must undergo in order to fulfill their function in host defense and tissue homeostasis. In summary, our results further establish a pivotal role for glucose and its breakdown via glycolysis in the control of morphodynamic activity in LPS-stimulated macrophages. Based on findings in other cell systems, we consider it 606143-89-9 19653627″ title=View Abstract(s)”>PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19653627 likely that for exerting this role, the multitalented properties of glucose as metabolic precursor and molecule for use in post-translational modification of cytoskeletal (associated) structural proteins, recep- tors or signaling proteins are being used. More in depth investigation is required to further un