Figure 1. Insulin-induced tyrosine phosphorylation and expression level of IRS1 is altered by S668T and D674H mutants in 293T cells. One hundred μg of total protein was fractionated by 10% SDS-PAGE. Blots were first labeled with anti-phospho-specific antibodies, stripped and reprobed with non-phospho-specific antibody. Fold induction of phosphorylation was determined by dividing densitometric values of the phospho- band by that of total protein bands. Relative abundance of IRS1 was determined by dividing the densitometric values of IRS1 to that of beta-actin.
