FIG. 9.

Sustained activation in insulin signaling in Nfatc2^−/− Nfatc4^−/− mice is not cell autonomous. Primary hepatocytes isolated from Nfatc2^−/− Nfatc4^−/− (DKO) and control mice were challenged with insulin in vitro for times indicated (A). Cell extracts prepared were used to determine activation of insulin downstream effectors, including Akt and S6K protein kinases, by immunoblotting analysis using phospho-Akt (P-Akt) and phospho-S6K (P-S6K) antibodies. Expression of total Akt and S6K was used as the control. Activation of insulin signaling in primary macrophages isolated from peritoneal cavities of Nfatc2^−/− Nfatc4^−/− (DKO) or control mice was also determined (B).