Figure 3From: Integrin-mediated internalization of Staphylococcus aureus does not require vinculinTransient vinculin overexpression in 293 cells does not interfere with bacterial uptake. (A) 293 cells were transfected with constructs encoding GFP or GFP-vinculin. 48 h post-transfection whole cell lysates (WCLs) were prepared and analyzed for vinculin and β-tubulin expression by immunoblotting. Representative blots are shown. (B) Cells transfected as in (A) were compared for integrin α5 surface expression by flow cytometry. Shown is a representative histogram of cells stained with α-integrin α5 antibody and a rhodamine-coupled secondary antibody (blue – GFP; red – GFP-vinculin) or cells stained with the secondary antibody only (grey lines). (C and D) Cells as in (A) were infected with S. aureus and S. carnosus for 2 h (MOI 20). Total cell associated or recovered viable intracellular bacteria were determined by bacterial cell adhesion (C) or gentamicin protection assays (D), respectively. Values are means ± SEM from 2 independent experiments done in quadruplicate, n=8. Mann-Whitney test was applied for statistical significance testing.Back to article page