Figure 7.

Effect of cixutumumab (A12) on the IGF1-stimulated INSR, IGF1R, AKT, and ERK phosphorylations in P69 cells. P69 cells were incubated with the A12 antibody in the presence or absence of IGF1. Levels of phosphorylated and total proteins were measured by western blot analysis. For phospho-receptor measurement, cell extracts were immunoprecipitated with anti-INSR (A) or IGF1R (B), electrophoresed, and immunoblotted with anti-phospho-INSR/IGF1R. AKT (E) and ERK1/2 (F) phosphorylation levels were measured using specific anti-phospho antibodies. Results of a typical experiment, repeated three times with similar results, are shown in the figure. Scanning densitometric analysis of the effect of A12 on INSR (C), IGF1R (D), AKT (G), and ERK1/2 (H) activation. Bars represent phospho-protein values normalized to the corresponding total protein levels. Bars are means±s.e.m. of three independent experiments. *P<0.05 vs control cells (G and H) and *P<0.05 vs A12+IGF1-treated cells (C and D).