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Figure 5 | Journal of Hematology & Oncology

Figure 5

From: TFPIα and TFPIβ are expressed at the surface of breast cancer cells and inhibit TF-FVIIa activity

Figure 5

Cell surface anticoagulant activity of PI-PLC treated cells. Confluent Sum102, MDA-MB-231, and HCAEC cells grown in 24-wells trays were treated with SFM (untreated), PI-PLC, monoclonal anti-TFPI antibody, or anti-TF antibody for 2 hours before the supernatant was removed and cells were washed and assayed for TF activity. Sum102 (A), MDA-MB-231 (B), and HCAEC (C) cells were incubated with FVIIa and FX for 1 hour before the reaction was stopped and a substrate to FXa was added. The results were analyzed colorimetrically at 405 nm. The amount of FXa generated is displayed as mean mU/mL + SD (n ≥ 7) of three to four independent experiments. Statistical comparisons were conducted between PI-PLC treated samples and untreated controls. (D) Fluorescence images of SFM + Glycerol (untreated, upper panels) and PI-PLC (bottom panels) treated Sum102 cells stained with antibodies against both isoforms of TFPI (green, left panels) and TF (red, middle-left panels). After merging the images, co-localization of the two antibodies was indicated by a yellow color (middle-right panels). Nuclear staining with DAPI is depicted in the left panels. One representative experiment of three is shown.

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