Thrombin increases OSM production and expression in human monocyte-derived macrophages. (A-B) MDMs were incubated with α-thrombin obtained from Sigma-Aldrich (A: □; B: ○) and α-thrombin obtained from Haematologic Technologies (A: ▩; B: ) at indicated concentrations or for indicated time periods in the absence (B: ●) or presence of α-thrombin (1 U/mL). Conditioned media of such treated cells were collected and OSM antigen was determined as described in “Quantification of OSM.” Values are given in nanograms per milliliter and represent mean values ± SD of 3 independent determinations. Experiments were performed 3 times with MDMs obtained from 3 different donors with similar results. A representative experiment is shown. (C) MDMs were incubated with or without α-thrombin obtained from Sigma-Aldrich (□) and α-thrombin obtained from Haematologic Technologies (▩; 1 U/mL) for the indicated time period. mRNA was prepared and analyzed by RealTime-PCR with specific primers for OSM and GAPDH as described in “Real-time PCR.” OSM mRNA levels were normalized according to the respective GAPDH mRNA levels. Values are given as x-fold of control, which was set as 100%, and represent mean values ± SD of 3 independent determinations. Experiments were performed 2 times with MDMs obtained from 2 different donors with similar results. A representative experiment is shown. *P < .001 (α-thrombin; Sigma-Aldrich); §P < .001 (α-thrombin; Haematologic Technologies).