1. Volume 129, Issue 2, Pages 197-203 (February 2012)
Anthracycline treatment of the human monocytic leukemia cell line THP-1 increases phosphatidylserine exposure and tissue factor activity 
Jeremiah C. Boles, Julie C. Williams, Rachel M. Hollingsworth, Jian-Guo Wang, Sam L. Glover, A. Phillip Owens, David A. Barcel, Raj S. Kasthuri, Nigel S. Key, Nigel Mackman 
Thrombosis Research 
Volume 129, Issue 2, Pages (February 2012)
DOI: /j.thromres
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2. Fig. 1
Anthracyclines Concentration Dependently Increase Cellular TF Activity in THP-1 Cells: THP-1 cells were treated with cytotoxic chemotherapeutic agents for 24hours and were compared to untreated cells. A. THP-1 cells treated with increasing concentrations of doxorubicin (DOX). B. THP-1 cells treated with cytarabine (CYT). C. THP-1 cells treated with daunorubicin (DAUN). D. Time course of 0–24hours of THP-1 cellular TF activity after 1μg/ml DAUN treatment. * denotes statistical significance of P<0.05 compared to control. ** denotes statistical significance of P<0.001 compared to time 0.
Thrombosis Research  , DOI: ( /j.thromres )
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3. Fig. 2
Treatment of THP-1 Cells with DAUN does not Increase TF mRNA or Protein Expression: A. THP-1 cells treated with DAUN (1μg/ml) for 2hours were analyzed for TF mRNA expression using real time PCR. B. THP-1 cells were treated for 24hours with DAUN (1μg/ml) and then assayed for cellular TF protein using an ELISA. No significant differences were observed in TF mRNA or TF protein levels compared to control.
Thrombosis Research  , DOI: ( /j.thromres )
Copyright © 2011 Elsevier Ltd Terms and Conditions

4. Fig. 3
DAUN Concentration Dependently Increases MP TF Activity in the Culture Supernatant without an Increase in TF Protein Expression on the MPs: A. Levels of MP TF activity in the culture supernatant from THP-1 cells treated with increasing concentrations of DAUN. A concentration-dependent increase in MP TF activity was observed. B. MPs isolated from treated THP-1 cells were analyzed for TF protein expression by flow cytometry using a FITC labeled mouse anti-human monoclonal antibody versus an IgG isotype control. Samples were run with the same flow rate and events were collected for 30seconds. No significant increase in TF protein expression as determined by MFI was noted at any of the DAUN concentrations used. * indicates statistical significance (P<0.05) compared to control.
Thrombosis Research  , DOI: ( /j.thromres )
Copyright © 2011 Elsevier Ltd Terms and Conditions

5. Fig. 4
Increased MP TF Activity is Secondary to a Concentration Dependent Increase in the Number of PS+ MPs Released into the Culture Supernatant: A. Culture supernatant from untreated and treated THP-1 cells was collected and analyzed for MP PS content. B. MPs isolated from untreated and treated THP-1 cells were analyzed using flow cytometry for levels of PS using APC conjugated annexin V. Samples were run with the same flow rate and events were collected for 30seconds. No significant increase in PS expression as determined by MFI was noted at any of the DAUN concentrations used. C. MPs isolated from untreated and treated THP-1 cells were stained with calcein-AM and subsequently analyzed by flow cytometry. Events were considered to represent a MP if they fell within the mega-mix determined FSC and SSC gate as established by the ISTH SSC guidelines and were calcein-AM positive. D. MPs isolated from treated THP-1 cells were stained with both APC labeled annexin V and calcein-AM. * indicates statistical significance (P<0.05) compared to control.
Thrombosis Research  , DOI: ( /j.thromres )
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6. Fig. 5
DAUN Induces a Concentration Dependent Increase in Apoptosis and PS Exposure in THP-1 cells: A. Unteated and DAUN treated THP-1 cells were assessed for cell viability using Alamar Blue. Cells were also incubated with ZVAD (50μM) for 2hours prior to treatment with DAUN. B. Treated THP-1 cells were assessed for cell viability with a flow cytometry assay utilizing FITC labeled annexin V and the nuclear dye 7AAD. Cells were defined as viable if they were dual negative. C. THP-1 cells treated with increasing concentrations of DAUN were isolated and stained with APC labeled annexin V and analyzed by flow cytometry for PS MFI. D. THP-1 cells were treated with DAUN (1μg/ml) for 0–24hours and then stained with APC-labeled annexin V and analyzed by flow cytometry for PS MFI. * indicates statistical significance (P<0.05) compared to control or time 0. ** indicates statistical significance (P<0.05) compared with THP-1 cells treated with DAUN (1μg/ml). *** indicates statistical significance (P<0.001) compared to untreated control cells, time 0.
Thrombosis Research  , DOI: ( /j.thromres )
Copyright © 2011 Elsevier Ltd Terms and Conditions

7. Fig. 6
Annexin V and Lactadherin Abolish the Increase in Cellular TF Activity in THP-1 cells treated with DAUN: THP-1 cells treated with DAUN (1μg/ml) were incubated with either annexin V 100μM (DAUN+Annexin V) or lactadherin 100 uM (DAUN+Lactadherin) for 15minutes prior to the addition of FX and FVIIa and levels of FXa were determined. * indicates statistical significance (P<0.05) compared to control. ** indicates statistical significance (P<0.05) compared to DAUN treated THP-1 cells.
Thrombosis Research  , DOI: ( /j.thromres )
Copyright © 2011 Elsevier Ltd Terms and Conditions