Supplementary file 1_A defined isothiocyanate mixture suppresses platelet signaling and metabolism to limit thrombosis in vivo.docx

The ITC mixture was administered before or after the carrageenan challenge. The ITC mixture inhibited platelet activation and aggregation induced by multiple agonists. It reduced peptide phosphorylation across multiple platelet signaling pathways, reduced intracellular calcium mobilization and the activation-associated oxidant signal, and impaired mitochondrial function.

When the ITC mixture was added shortly after activation, aggregometry signals consistent with destabilization of early platelet aggregates were observed whereas granule secretion was irreversible once initiated. In vivo, ITC mixture administration before or after the carrageenan challenge reduced thrombus formation and tissue injury, prolonged bleeding times, and reduced tail-necrosis length to a similar extent as the heparin reference group in this model.

These findings identify the ITC mixture as a modulator of platelet signaling and metabolism, provide mechanistic insight into ITC-mediated antithrombotic effects, and support further evaluation of ITC-based interventions in thrombotic disease.</p>