abolism and the extent of morphodynamic change that macrophages undergo. To apprehend the coupling between actin cytoskeletal remodeling and metabolic state we here investigated the ability of LPSstimulated (M1) RAW 264.7 and Maf-DKO macrophages to maintain functional activity under conditions where they were forced to shift between Aphrodine chemical information PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19652232 a (anaerobic) glycolytic or oxidative metabolism. RAW 264.7 and Maf-DKO cell lines were chosen because they are in vitro manipulable models that have retained marked plasticity to stimulus-directed polarized activation, but lack the phenotypic heterogeneity that is characteristic for primary macrophages [45,46]. We report on a stringent dependency of morphodynamics of LPS-stimulated macrophages upon sufficient glucose supply. Cell Culture RAW 264.7 cells (kind gift from Dr. Hong-Hee Kim, Department of Cell and Developmental Biology, School of Dentistry, Seoul National University, Korea; [47]) were maintained in high-glucose DMEM (Gibco, Life Technologies, Paisley, UK) supplemented with 10% heat inactivated FBS (PAA laboratories, Pasching, Austria), 1 mM sodium pyruvate, and 4 mMGlutaMAX (Gibco, Life Technologies, Paisley, UK), at 37uC in a humidified atmosphere with 7.5% CO2. Maf-DKO cells (kind gift from Dr. Michael H. Sieweke, Centre d’Immunologie de Marseille-Luminy (CIML), Universite Aix-Marseille, France; [46]) � were maintained in the same way except that medium was supplemented with 20% conditioned medium from L929-cells containing macrophage colony stimulating factor (M-CSF). DNA Constructs and Transfection Plasmid pEYFP-N1-DATG-Lifeact was constructed as follows: Lifeact [48] cDNA, containing human codon sequences flanked by a 59 BglII and 39 EcoRI restriction site, was commercially s

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