Fig. 7. Re-expression of full-length DG or DG mutants in a completely DG-/- cell line restored laminin-111 binding on monolayer cell surfaces. (A) Western blot of cell extracts (10 µg protein) prepared from DG+/+ cells and from a DG-/- cell line (derived from MEpG cells) infected with retroviral vector (VEC) or that encoding full-length DG (wtDG) or various ß-DG cytoplasmic deletions (DEL A, B and C). Blots were incubated with antibodies specific for
-DG, N-terminal ß-DG (right panel), C-terminal ß-DG (left panel), or E-cadherin (loading control), followed by HRP-conjugated secondary antibodies. Sizes of molecular mass markers are shown in kDa. (B) Paired immunofluorescent images of cells in A, co-stained for
-DG and nuclei, using FITC-labeled secondary antibody and propidium iodide, respectively. (C) Immunofluorescent images of cells in A, treated for 4 hours with 10 nM exogenously added laminin-111-FITC. Corresponding phase images are shown in the bottom panel. Bars, 10 µm.