Figure 6.

AnxA2 antibody inhibits the EGF-induced AKT pathway in MDA-MB-231 and JIMT-1 cells. (A) MDA-MB-231 and JIMT-1 cells after 70% confluence were cultured overnight in serum free medium, and then incubated with EGF (50 ng ml⁻¹) for 20 min after 2 h of heat inactivated AnxA2 (D1/274.5) antibody (2 μg ml⁻¹) AnxA2 (D1/274.5) antibody (2 μg ml⁻¹) pretreatment. Western blot analyses were carried out with indicated antibodies. (B) A bar graph showing densitometric analysis of bands used in (A). Each bar represents the mean±s.e. of three independent experiments. (C) After 12 h serum starvation, MDA-MB-231 and JIMT-1 cells were treated with or without EGF (50 ng ml⁻¹) for 20 min after 2 h of PI3K inhibitor (10 μg ml⁻¹) pretreatment. The cell lysate was analysed for p-PDK1 (Ser-241), p-AKT (Ser-473) by western blotting.