Fig. 5
PDGF-AB enhances uPAR/P-FAK interactions in migrating BM-MSC. a uPAR (green) and P-FAK (red) repartition in migrating cells. BM-MSC were seeded in a Labtek chamber coated with type I dermal collagen. After the scratch was performed, cells were grown in serum-free control medium or treated with PDGF-AB for 3 hours. uPAR and P-FAK stainings were merged to show colocalization (yellow). Nuclei were stained with DAPI. Scale bars: 10 μm. b uPAR–P-FAK coimmunoprecipitation. Two hours after adherence on type I dermal collagen, cells were seeded in serum-free control medium (−) or treated with PDGF-AB (+) for 1 hour. uPAR was immunoprecipitated from cell lysates with anti-uPAR monoclonal antibodies or isotypic control IgG1. Immunoprecipitates were analyzed by SDS-PAGE and blotted for uPAR and P-FAK. (Left) Western blot representative of three experiments. T0, lysate of cells before seeding; Adh., lysate of cells allowed to adhere for 2 hours on type I dermal collagen. (Right) P-FAK blots quantification by densitometry analysis using ImageJ software. Data expressed as the P-FAK fold change after PDGF-AB treatment compared with serum-free control medium from three independent experiments (one donor per experiment). PDGF-AB platelet-derived growth factor AB, P-FAK autophosphorylated focal adhesion tyrosine kinase, uPAR urokinase-type plasminogen activator receptor