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. 2015 Oct 17;4(12):951–960. doi: 10.1016/j.molmet.2015.09.013

Figure 6.

Figure 6

TRAF3 negatively regulates insulin signaling in the liver. A.TRAF3f/f and HKO male mice (7–8 weeks) were fed a HFD for 10–12 weeks, fasted for 20–24 h, and injected with insulin (2 units/kg body weight) via inferior vena. Liver extracts were prepared 5 min after insulin stimulation and immunoblotted with antibodies against phospho-Akt (pThr308 and pSer473) or Akt. Insulin-stimulated phosphorylation of Akt was quantified and normalized to total Akt levels. B. DKO and ob/ob;TRAF3f/f male mice (10 weeks) were fasted overnighted and stimulated with insulin (4 units/kg) for 5 min. Liver extracts were immunoblotted with antibodies against phospho-Akt (pSer473) or Akt. Phosphorylation of Akt was quantified and normalized to total Akt levels. C. Primary hepatocytes were prepared from TRAF3f/f and HKO male mice (8–9 weeks) and stimulated with 100 nM insulin for the indicated time. Cell extracts were immunoblotted with antibodies against phospho-Akt (pSer473) or Akt. *p < 0.05.