Figure 3. Insulin-induced phosphorylation and expression level of ERK kinase is altered by S668T and D674H mutants in 293T cells. One hundred μg of total protein was fractionated by 10% SDS-PAGE. Blots were 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 to that of total protein bands. Relative abundance of ERKs was determined by dividing the densitometric values of ERKs to that of beta-actin.
