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HEMCAM/CD146 downregulates cell surface expression of b1 integrins

Sandrine Alais^1, Nathalie Allioli^1, Cristina Pujades^1, Jean-Loup Duband1, Olli Vainio^2, Beat A. Imhof³ and Dominique Dunon¹

¹UMR-CNRS 7622, Université Pierre et Marie Curie, 75005 Paris, France
²Department of Medical Microbiology, Turku University, FIN-20520, Finland
³Department of Pathology, University of Geneva, CH-1211, Geneva, Switzerland

Cells were grown at confluence and cell monolayers were wounded by a yellow tip. Adherent cells were washed twice and incubated in complete L15 medium (SVF, 10%; penicillin, 10 UI/ml; streptomycin, 100 mg/ml) under an inverted microscope in a temperature controlled room. Culture behaviors were filmed for 5 hours. (A) Mock-transfected cells. Real time=5 hours; interval=7 minutes. (B) HEMCAM-s transfected cells. Real time=5 hours; interval=15 minutes.

Motile activity was studied by videomicroscopy 24 hours after plating the cells in culture dished plates. Mock- and HEMCAM-transfected cells were motile but the structure of protrusions was different between cell lines. Mock-transfected cells (A) exhibit long and thin filopodia, whereas HEMCAM-transfected cells (B) gave rise to short and puffy protrusions. These experiments suggested that these alterations of motility were involved in the inhibition of migration. Adherent cells were washed twice and incubated in complete L15 medium (SVF, 10%; penicillin, 10 UI/ml; streptomycin, 100 mg/ml) under an inverted microscope in a temperature-controlled room. Mock-motility corresponds to mock-transfected cells; HEMCAM-motility corresponds to HEMCAM-transfected cells.

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