The recent findings that NFAT proteins have a critical role in the adipocyte differentiation process and in the regulation of Fostamatinib (R78 cool in newly differentiated adipocytes prompted us to look for the regulatory involvement of NFAT proteins in other biological and metabolic processes of mature adipocytes. To determine a functional role of NFAT in mature adipocytes, we studied 3T3-L1 adipocytes that had been cultured for >15 days after induction of differentiation. Such long-term cultures of differentiated adipocytes are representative of mature adipocytes, given that expression of proteins linked to adipocyte differentiation does not change significantly after 8 days of such treatment [14] and [15]. We utilized CsA and GSK3β inhibitors to look for changes in physiologic processes and expression of genes relevant to mature adipocyte lipolysis and metabolism.
Materials and methods
CsA was from Sigma (St. Louis, MO) and 4-benzyl-2-methyl-1,2,4-thiadiazolidine-3,5-dione (TDZD- cool was from Calbiochem (La Jolla, CA). Recombinant murine tumor necrosis factor-α (TNFα) was prepared as previously described [16]. BSA was from Intergen (Purchase, NY).
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Statistical analysis Statistical analysis was performed using GraphPad Prism Software
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