Figure 3. Unimpaired insulin induction of GCK and SREBF1 mRNAs in hepatocytes overexpressing TRB3/NIPK.

The method for hepatocyte culture and transduction was as described in Figure 2, except that cells were harvested for isolation of total RNA after 7 h of insulin stimulation. Both uninfected hepatocytes and hepatocytes transduced with an AdEasy-1 lacZ vector were used as controls. Agarose gel electrophoresis was performed with 16 μg of RNA per lane. Northern blotting was done on a nylon membrane hybridized sequentially to ³²P-labelled cDNA probes for GCK, SREBF1 and ribosomal protein S26. Positions of 28 and 18 S rRNAs are shown on the right.