Files in this Data Supplement:

• Figure I - (TIF) (8.03 MB). Lower concentrations of dipyridamole attenuate, but do not eliminate, MCP-1 synthesis. Platelets and monocytes were stimulated with thrombin (0.1 U/mL) for 18 hours in the presence of vehicle, dipyridamole (Dip, 2 μg/mL), aspirin (ASA, 250 ng/mL), or dipyridamole plus aspirin (2 μg/mL of Dip plus 250 ng/mL of ASA). MCP-1 generated by the monocytes was determined for each condition. The bars represent the mean±SE for 6 independent experiments.
• Figure II - (TIF) (22.7 MB). Dipyridamole does not significantly reduce IL-8 synthesis. Platelets and monocytes were stimulated with thrombin (0.1 U/mL) for 4 (A) or 18 (B) hours in the presence of vehicle, dipyridamole (Dip, 5 μg/mL), aspirin (ASA, 625 ng/mL), or dipyridamole plus aspirin (5 μg/mL of Dip plus 625 ng/mL of ASA). IL-8 generated by the monocytes was determined for each condition and time point. The bars represent the mean±SE for 6 independent experiments.
• Figure III - (TIF) (15.7 MB). Dipyridamole does not block COX-2 expression in platelet–monocyte aggregates. COX-2 protein expression was measured in platelets, monocytes, or platelet–monocyte suspensions that were pretreated with the specific drugs and subsequently stimulated with thrombin (Thr, 0.1 U/mL) for 18 hours (top panel). Corresponding densitometry data for this western blot analysis are depicted in the bottom panel. Here, the data are expressed as a fold change of COX-2 expression in the vehicle-treated group. This Western blot is representative of 5 independent experiments.
• Figure IV - (TIF) (11.7 MB). Dipyridamole reduces PDE4A phosphorylation in thrombin-activated platelets. Platelets were pretreated with dipyridamole (5 μg/mL) and left quiescent or stimulated with thrombin (0.1 U/mL) for the designated time periods. Proteins were separated by SDS-PAGE, and PDE4A phosphorylation was measured using a phosphospecific antibody (see Methods). This antibody recognizes several PDE4A long form variants (molecular weights between 102 and 109 kDa) that include PDE4A5, PDE4A8, and PDE4Ax (see brackets on right side of gel). The antibody also reacts with another band at ~76 kDa (see arrow on right side of gel) that is presumably another isoform of PDE4A that remains uncharacterized. This gel is representative of 3 independent experiments.
• Figure V - (TIF) (10.3 MB). Exogenous adenosine deaminase (ADA) reverses that inhibitory effect of dipyridamole on LPS-induced MCP-1 gene expression. Monocytes were pretreated with dipyridamole (DIP) in the presence or absence of ADA (1.0 U/mL). The cells were subsequently stimulated with LPS as described in Methods, and MCP-1 protein levels were measured by ELISA. The data are expressed as a percentage of LPS-induced MCP-1 synthesis and the bars in the graph represent the mean±SEM of 5 experiments. The asterisk (*) in this figure represents P‹0.05 versus LPS-stimulated cells or LPS-stimulated cells that were pretreated with DIP and ADA.