Fig. 4. Increased levels of activated 1 integrins but lower overall levels of 1 integrins are expressed on the plasma membrane of Ehd1^–/– MEF cells. MEF cells grown on uncoated (A-C) or fibronectin-coated (A; FN) plates were harvested by brief trypsinization (A; Trypsin and B,C) or scraping (A; Scrape), fixed and subjected to flow cytometry analysis under non-permeabilizing conditions with secondary antibody only (A-C) or first with 9EG7 antibodies that preferentially recognize ligand-bound 1 integrins (BD Pharmingen) (A), conformation-independent 1 integrin antibodies (MB1.2, Chemicon) (B), anti-LAMP1 antibodies (C) or antibodies directed against the epidermal growth factor (EGF) receptor (C) prior to secondary antibodies. The graphs depicted are representative of 4-6 independent experiments. +/+, Ehd1^+/+ MEF cells; –/–, Ehd1^–/– MEF cells.