Confluent endothelial monolayers were treated with increasing peptide concentrations in HBSS (1 h at 4 °C, unless otherwise stated). After removing unbound peptide, cells were either incubated in appropriate cell culture medium (37 °C) for varying time-points, or directly treated with neutravidin-FITC or albumin-FITC (0.83 mM, in HBSS, 30 min at 4 °C). After removing unbound protein with thorough HBSS washes, fluorescence (490/525 em/ex) was quantified with a Spark multimode microplate reader (Tecan) or imaged through confocal microscopy (Additional file 1: Diagram S1a–c).
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