HUVEC cells were pelleted by centrifugation at 2000 rpm for 5 min before harvesting the conditioned medium. sVEGFR1 levels were measured in a Luminex assay, using the human anti-VEGFR1 antibody (clone 49560, R&D Systems). Measurements were performed using an in-house developed and validated (ISO9001 certified) multiplex immunoassay based on Luminex xMAP technology (Luminex, Austin TX USA) as described (26).

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