Figure 4

Met and EGFR in A431 cells are independently activated upon cell adhesion. (a) An equal amount of whole cell lysates from attached (att), suspended (sus), and replated (rep) A431 cells in the presence (+) or absence (-) of 0.5 μM PHA665752 (a specific inhibitor for Met) for 60 min was analysed by immunoblotting with antibodies as indicated. (b) The control A431 cells and those stably expressing shRNAs specific to Met (sh-Met) or luciferase (sh-Luc) were kept in suspension for 30 min and replated on dishes coated with collagen for 60 min. An equal amount of whole cell lysates from attached (att), suspended (sus), and replated (rep) cells was analysed by immunoblotting with antibodies as indicated. (c) The A431 cells stably expressing shRNAs specific to EGFR (sh-EGFR clone #1 and #2) or luciferase (sh-Luc) were suspended in serum-free medium for 30 min and then replated on dishes coated with collagen for 60 min. An equal amount of whole cell lysates from attached (att), suspended (sus), and replated (rep) cells was analysed by immunoblotting with antibodies as indicated.