Hcpg 0130

The day before each experiment, near-confluent tenocytes were trypsinized for 10 minutes in 0.25% Trypsin-EDTA in a humidified incubator (5% CO₂, 37°C). Cells were then washed in culture media, centrifuged, strained to ensure a single cell suspension, and counted using a hemocytometer. The cells were then pelleted, suspended in control media (MEM α supplemented with 1% FBS and 1% Pen Strep), seeded onto the collagen-coated plates (70,000 cells/2 mL media/well) and incubated overnight in control media (MEM α and 1% FBS). The next day, time 0 samples were collected, and the remaining samples were treated with fresh control media with or without 10 ng/mL of TGF-β1 (R&D Systems, #240-B-010), 20 μg/mL of human glu-plasminogen (Haematologic Technologies, HCPG-0130), 50 ng/mL of tPA (Technoclone, #TC41072), 5 μg/mL of tranexamic acid (Sigma-Aldrich, #857653) or their respective vehicles (specified by the manufacturers). After 48 h, cells and media samples were collected and immediately stored at −80°C for later assessment of gene, protein, and protease activity levels. Four separate experiments (n=3 per experiment) were performed, but the number of samples used for each assay varied and is indicated where appropriate.