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SKOV3 and HEY cells (2 × 104) were seeded in a 96-well culture plate precoated with 50 μl/well of growth factor reduced Cultrex (Trevigen) and stimulated with ET-1 or MAC for 16 h. Upon treatment, tubule-like structure formations were visualized with an inverted microscope with a 20× magnification. Representative images were captured with a ZOE Fluorescent Cell Imager (BioRad Laboratories). Tube formation was analyzed by using Angiogenesis Analyzer for ImageJ (NIH) measuring the number of nodes and the tube length.
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