Materials and Methods
Materials
Thrombin was obtained from Sigma Aldrich (St Louis, MO). ML161 (PAR-1 antagonist) was purchased from TargetMol (Boston, USA). The anti-NF-κB p65, anti-phospho-NF-κB p65, anti-IκBα, anti-phospho-IκBα, anti-Erk, anti-phospho-Erk, anti-STAT3, anti-phospho-STAT3, anti-Akt, anti-phospho-Akt were purchased from Cell Signaling Technology (Danvers, MA). The antibody specific for thrombin, PAR-1, and CD31 were from Santa Cruz (Santa Cruz Biotechnology, Santa Cruz, CA). The anti-MMP9, anti-IL6 and anti-GAPDH were from Proteintech (Proteintech Group, Chicago, IL). The kit for monitoring the activation of RhoA was obtained from Cytoskeleton (Denver, CO). r-hirudin and DTIP were obtained from our group, and expression and purification of r-hirudin and DTIP has been described previously (Mo, Zhang, Chen, Wang & Song, 2009; Zhao et al., 2017).