Significance Activated protein C (aPC) is a promising therapeutic and exhibits multiple pharmacological benefits in preclinical studies, including in sepsis. aPC activates protease-activated receptor-1 (PAR1) and signals via β-arrestin-2 to promote endothelial barrier stabilization and anti-inflammatory and anti-apoptotic activities. The present study demonstrates that different aPC/PAR1 cytoprotective responses are uniquely regulated by discrete β-arrestin-2–mediated signaling pathways that are modulated by coreceptors. Thus, β-arrestin-2 functions as a central driver of distinct, aPC/PAR1-induced cytoprotective signaling pathways to control barrier protection and anti-apoptotic activities.