Figure 1. Mig-6 inhibits EGF-mediated cell migration and filopodia formation.

H1299 cells were stably infected with lentivirus expressing GFP and Myc-tagged Mig-6 (Mig-6), or GFP alone (VEC). (A) Whole-cell lysates from these stable cells were subjected to immunoblotting, as indicated. (B) Migration of these stable cells was assessed by wound-healing assay in the presence or absence of EGF (100 ng/mL). Scale bars = 200 μm. (C) Wound closure data was quantitated and presented as means and SE from three independent experiments. *P < 0.05. (D) Stable H1299 cells were subjected to cell migration assays using transwell systems in the presence of EGF. Migratory cells were fixed, stained with crystal violet, and photographed under a light microscope. Representative pictures are shown. Scale bars = 100 μm. (E) Results from migration assays were quantitated and presented as means and SE from three independent experiments. ***P < 0.001. (F) Stable H1299 cells were grown in the presence of EGF for 24 hours. Cells exhibiting filopodia were visualized by fluorescent microscopy. F-actin was visualized with phalloidin (red), and nuclei was counter-stained with DAPI (blue). Note that Mig6 inhibited EGF-induced filopodia formation. Representative fluorescent microscopic images and insets are shown. Scale bars = 50 μm. (G) Percentage of cells with multiple filopodia (defined as cells exhibiting at least 20 filopodia) were quantified and presented as means and SE from three independent experiments performed in duplicate. *P < 0.05.