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. 2009 Nov 10;101(12):2023–2029. doi: 10.1038/sj.bjc.6605406

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Copyright © 2009 Cancer Research UK

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Secretion of MPs with TF-fVIIa activity by ovarian cancer cells. (A) fXa generation activity of the supernatant and precipitate fraction of conditioned media prepared from OVSAYO and OVISE cells cultured under 1% O₂. Procoagulant activity predominantly presented in the precipitate fraction and this activity was completely blocked by anti-TF (5G9) antibody treatment. ^*P<0.001, ^**P=0.001. (B) Flow cytometry analysis of precipitate fraction immunostained with PE-coupled anti-TF antibody and/or FITC-coupled annexin V showed that precipitate fraction prepared from conditioned media of OVSAYO cells was annexin-V positive. (C) The extracellular procoagulants analysed by flow cytometry. The grey peak corresponds to the 1-μm calibration microspheres. The white peak corresponds to the precipitate fraction prepared from conditioned media of OVSAYO cells. (D) Flow cytometry analysis of the precipitate fractions prepared from serum-free conditioned media of OVSAYO cells cultured under normoxia or 1% O₂ for 24 h. (Da) The MP fraction prepared was TF positive. Background fluorescence was set with a fluorescent mouse IgG control. (Db) Dot plot of MP fraction immunostained with PE-coupled anti-TF antibody and FITC-coupled annexin V. Percentages shown in the upper right segment of the figure denote the relative amount of TF-positive MPs. (E) Flow cytometry analysis of the precipitate fractions prepared from serum-free conditioned media of OVSAYO cells cultured under normoxia or 1% O₂ for 24 h. (Ea) The MP fraction prepared was fVII positive. Background fluorescence was set with a fluorescent mouse IgG control. (Eb) Dot plot of MP fraction immunostained with PE-coupled anti-TF antibody and FITC-coupled anti-fVII antibody. Percentages in the upper right segment of the figure denote amounts of fVII-positive fractions relative to total MPs.