In Vivo Ferric Chloride Carotid Artery Injury–Induced Thrombosis Model

C57BL/6 mice were exposed as discussed above, and the assay was performed as we described before.38, 39, 40 Briefly, mice were anesthetized with Avertin (2.5%), and the left carotid artery was exposed and cleaned with normal saline (37°C), before baseline carotid artery blood flow was measured with Transonic Micro‐Flowprobe (Transonic Systems Inc, Ithaca, NY). After stabilizing blood flow, 7.5% ferric chloride was applied to a filter paper disc (1‐mm diameter) that was immediately placed on top of the artery for 3 minutes. Blood flow was continuously monitored for 20 minutes or until blood flow reached stable occlusion (no blood flow for 2 minutes). Data were recorded, and time to vessel occlusion was calculated as the difference in time between stable occlusion and removal of the filter paper (with ferric chloride). An occlusion time of 20 minutes was considered as the cutoff time for statistical analysis.