Figure 5
Vitronectin oligomers induce membrane leakage. Large unilamellar vesicles (100 nm in diameter) loaded with the fluorophore-quencher pair ANTS-DPX were incubated with indicated concentrations of protein. Recombinant vitronectin oligomers incubated with 500 μM LUVs resulted in appreciable leakage of vesicles in a dose-dependent fashion (grey bars) as compared to recombinant vitronectin freshly dissolved in phosphate buffer (white bars). Membrane leakage was assessed by monitoring ANTS fluorescence (λex = 353 nm, λem = 520 nm). Intensities were normalized to the intrinsic fluorescence of the vesicles and percent leakage was estimated by dividing the sample and maximum fluorescence intensity values. Data shown are the combined average of three independent experiments. Error bars represent the standard error of the mean.