The novel tethered ligand TR47 peptide induces PAR1-dependent signaling in endothelial cells. Phosphorylation of pSer473-Akt and pSer9-GSK3β by TR47 peptide was monitored in endothelial cells. (A) Time course of Akt phosphorylation at Ser473 by TR47 (50μM). (B) Time course of GSK3β phosphorylation at Ser9 by TR47 (50μM). (C) Phosphorylation of Akt at Ser473 by TR47 (50μM) or a control scrTR47 peptide (50μM) over time. (D) Phosphorylation of GSK3β at Ser9 by TR47 (50μM) and a control scrTR47 peptide (50μM) over time. (E) TR47 peptide dose dependence for phosphorylation of Akt at Ser473 at 60 minutes. (F) Time course for phosphorylation of Akt at Ser473 by TR47 (50μM) in the presence of the PAR1 inhibitor, SCH79797, or vehicle control (DMSO). (G) Time course for phosphorylation of GSK3β at Ser9 by TR47 (50μM) in the presence of the PAR1 inhibitor, SCH79797, or vehicle control (DMSO). (A-B) Representative experiments. (C-G) Data points represent the mean ± SEM (N ≥ 3).