Immunoblot analysis of SREBP-1 and -2 in membranes and nuclear
extracts from livers of control rats and rats treated with STZ without
or with INS. Livers from the three groups of rats in Table 1 (four male
rats per group) were pooled, and aliquots of the membrane pellet (30
μg of protein) and nuclear extract (30 μg) were subjected to
SDS/PAGE (8% gel). Immunoblot analysis was performed with 5
μg/ml rabbit anti-rat SREBP-1 IgG (lanes 1, 2, and 3) or
anti-rat SREBP-2 IgG (lanes 4, 5, and 6) as the primary antibody and
0.25 μg/ml horseradish peroxidase-coupled donkey anti-rabbit
IgG as the secondary antibody. Filters were exposed to Reflection
NEF496 film for 15 s (lanes 1, 2, and 3) or 30 s (lanes 4, 5,
and 6) at room temperature. P and N
denote the precursor and cleaved nuclear forms of SREBP,
respectively.