Figure 3

Adhesion-induced activation of Met and EGFR does not rely on Src, ROS, or cytoskeleton. (a) The control A431 cells and those stably expressing shRNAs specific to Src (sh-Src) 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. (b) An equal amount of whole cell lysates from attached (att), suspended (sus), and replated (rep) A431 cells in the presence (+) or absence (-) of 30 mM NAC for 30 min was analysed by immunoblotting with antibodies as indicated. (c) An equal amount of whole cell lysates from attached (att), suspended (sus), or replated (rep) A431 cells in the presence (+) or absence (-) of 2 μM cytochalasins D (CD) for 60 min was analysed by immunoblotting with antibodies as indicated. The cells were stained for F-actin with phalloidin. Representative images for F-actin staining are shown. Scale bar, 20 μm. (d) An equal amount of whole cell lysates from attached (att), suspended (sus), or replated (rep) A431 cells in the presence (+) or absence (-) of 30 μg/ml nocodazole (NOC) for 60 min was analysed by immunoblotting with antibodies as indicated. The cells were stained for microtubules with anti-α-tubulin. Representative images are shown. Scale bar, 20 μm.