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NETs increase TF expression and procoagulant activity in breast cancer cell lines. MCF7 and T-47D cells were starved and stimulated with NETs (500 ng/mL) for 24 h. TF mRNA expression was evaluated by qRT-PCR, and GAPDH was used as a reference gene. The relative expression of mRNA was calculated using the ΔΔCT method ( A , D ). Flow cytometry was performed using PE-conjugated antibody <t>anti-CD142</t> ( B , E ). A clotting assay was carried out using platelet-poor plasma incubated with breast cancer cells. The reaction was initiated with CaCl 2 ( C , F ). Values represent the mean ± standard deviation of 3 independent experiments. Statistical analysis was performed using the unpaired t -test. n.s ., without significance and *** p -value < 0.001.
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NETs increase TF expression and procoagulant activity in breast cancer cell lines. MCF7 and T-47D cells were starved and stimulated with NETs (500 ng/mL) for 24 h. TF mRNA expression was evaluated by qRT-PCR, and GAPDH was used as a reference gene. The relative expression of mRNA was calculated using the ΔΔCT method ( A , D ). Flow cytometry was performed using PE-conjugated antibody <t>anti-CD142</t> ( B , E ). A clotting assay was carried out using platelet-poor plasma incubated with breast cancer cells. The reaction was initiated with CaCl 2 ( C , F ). Values represent the mean ± standard deviation of 3 independent experiments. Statistical analysis was performed using the unpaired t -test. n.s ., without significance and *** p -value < 0.001.
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NETs increase TF expression and procoagulant activity in breast cancer cell lines. MCF7 and T-47D cells were starved and stimulated with NETs (500 ng/mL) for 24 h. TF mRNA expression was evaluated by qRT-PCR, and GAPDH was used as a reference gene. The relative expression of mRNA was calculated using the ΔΔCT method ( A , D ). Flow cytometry was performed using PE-conjugated antibody anti-CD142 ( B , E ). A clotting assay was carried out using platelet-poor plasma incubated with breast cancer cells. The reaction was initiated with CaCl 2 ( C , F ). Values represent the mean ± standard deviation of 3 independent experiments. Statistical analysis was performed using the unpaired t -test. n.s ., without significance and *** p -value < 0.001.

Journal: Cancers

Article Title: TF/PAR2 Signaling Axis Supports the Protumor Effect of Neutrophil Extracellular Traps (NETs) on Human Breast Cancer Cells

doi: 10.3390/cancers16010005

Figure Lengend Snippet: NETs increase TF expression and procoagulant activity in breast cancer cell lines. MCF7 and T-47D cells were starved and stimulated with NETs (500 ng/mL) for 24 h. TF mRNA expression was evaluated by qRT-PCR, and GAPDH was used as a reference gene. The relative expression of mRNA was calculated using the ΔΔCT method ( A , D ). Flow cytometry was performed using PE-conjugated antibody anti-CD142 ( B , E ). A clotting assay was carried out using platelet-poor plasma incubated with breast cancer cells. The reaction was initiated with CaCl 2 ( C , F ). Values represent the mean ± standard deviation of 3 independent experiments. Statistical analysis was performed using the unpaired t -test. n.s ., without significance and *** p -value < 0.001.

Article Snippet: An amount of 1 × 10 6 cells/mL suspended in serum-free DMEM medium was washed twice with cold FACS buffer (PBS containing 0.01% sodium azide and 3% FBS) and labeled with PE-conjugated antibody anti-CD142 (TF) (BD Pharmingen, EUA) for 30 min at room temperature and fixed with 4% paraformaldehyde.

Techniques: Expressing, Activity Assay, Quantitative RT-PCR, Flow Cytometry, Coagulation, Incubation, Standard Deviation