Figure 7: CYP2E1 induces insulin resistance and ROS production in BI-1 knock-out primary hepatocytes.

Primary hepatocytes from BI-1 wild-type and BI-1 knock-out mice were treated with 250 μM palmitate in presence or absence of insulin. Cell lysates were immunoprecipitated with p-Tyrosine antibody and immunoblotted with antibodies against IRS1/2 (a). Hepatocytes from BI-1 knock-out mice were transiently transfected with non-specific-siRNA or CYP2E1 siRNA. Cells were treated with 250 μM palmitate in presence or absence of insulin. Cell lysates were immunoprecipitated with p-Tyrosine antibody and immunoblotted with antibodies against IRS1/2 (b). The Cell lysates were immunoblotted with anti-p-Akt, Akt, p-FoxO1, FoxO1, p-GSK3β, GSK3β or β–actin antibody (c). In non-specific-siRNA or CYP2E1siRNA-transfected BI-1 knock-out hepatocytes, 250 μM palmitate was treated for 12 h, then 100 μM DCF-DA was loaded for 30 min. After washing, fluorescence was measured by FACS analysis (d). Representative data performed in three separate measurements.