Fig. 7

Effect of MAP kinase pathway inhibition on the protection by IGF-1 of breast cancer cells from anoikis. MCF-7 cells were trypsinised, resuspended in serum-free medium alone or serum free medium and 10 ng/ml IGF-1, and in the absence or presence of 1 μM U0126 and then cultured for 24 h in 35-mm-diameter poly-HEMA-coated wells (a) or as attached cells (c). Cells were lysed and the amount of cleaved PARP, phosphorylated IGF receptors, phosphorylated ERK1 and ERK2, type I IGF receptor, ERK1 and ERK2 and GAPDH were measured by western transfer analysis in aliquots of 10 μg of protein. Representative images are shown with the images for each phosphorylated protein above those of the equivalent total protein (a). The means ± SEM from three experiments are shown (b). Statistical significance of differences between untreated cells and cells treated with IGF-1 (*), or between cells treated with IGF-1 alone and in the presence of the inhibitor (**) are indicated (p < 0.05; ANOVA)