The versatility of RSV lies in its diverse targeting of membrane and intracellular receptors, signaling mole cules, biogenesis enzymes, oxidative systems, DNA repair mechanisms and transcription factors, as well as in the wide range of possible RSV induced effect, including cellular VX-770 proliferation, cell Inhibitors,Modulators,Libraries cycle arrest, differ entiation and cell death. To elucidate the underlying mechanism of RSV ac tion, much research has been focused on different tis sues and cell types such as myocardial cells and hepatocytes. But, since RSV has been shown to act on skeletal muscle metabolism and function, less attention has been given to its effects on myogenesis. In vitro model for myogenesis study C2C12 murine immortalized cell line provides a good in vitro Inhibitors,Modulators,Libraries model for the study of the major steps of myo blasts proliferation and differentiation.
In this cellular model, undifferentiated myoblasts are recognizable as flat, fusiform or star shaped cells, which ap peared scattered on the substrate and rigorously mononu cleated. After reaching confluence Inhibitors,Modulators,Libraries or 24 hour after serum removal, C2C12 cells are considered myoblasts in an early differentiation stage and they are characterized by changes in myoblasts orientation, lengthening and thickening. Later, confluent mononucleated myocytes begin to fuse forming multinucleated myotubes, positive for the characteristic muscle specific protein MyHC. Myotubes become wider and longer over the next few days as additional myocytes fusion. Multinucleated and large myotubes appear to form a network with numerous nuclei arranged in multiple linear arrays.
In the present work we investigated potential mecha nisms mediating the effects of two different doses of Resveratrol on cell cycle regulation, skeletal muscle differentiation Inhibitors,Modulators,Libraries and during the genesis of hypertrophy in C2C12 myoblastic cells. Methods Materials Inhibitors,Modulators,Libraries Mouse C2C12 myoblastic cells were purchased from the European Collection of Animal Cell Cultures. Re agents were purchased from Sigma Chem. Primary antibodies anti MyoD, anti Myf 5, anti Akt12, anti MyHC, anti p21, anti Myogenin, anti Calnexin, anti GDF 8, anti IGF 1, anti N Cadherin, anti p120, anti AMPK12, anti pERK12, anti ERK1, anti ERK2, anti p53 monoclonal or polyclonal primary antibodies and the peroxidase conjugated or rhodamine conjugated secondary antibodies were purchased from Santa Cruz Biotechnology.
Alpha Sarcomeric Actinin pri mary antibody was purchased from Sigma Chem. Co. Anti phospho Akt and phospho AMPK were purchased from Cell Signaling Technology. In particular, Resveratrol was purchased from Sigma Chem. selleck bio and, according to the manufacturers instruction, it was dissolved in sterile water. Experimental procedures C2C12 cells were maintained at 37 C in humidified 5% CO2 atmosphere in a growth medium containing DMEM supplemented with 20% FBS, 1% penicillin streptomycin and 1% L glutamine up to 70% confluence.
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