In Vivo Thrombosis Model

This experiment was performed as previously described.^22,25,26 JUUL e-cigarette and clean air-exposed mice were anesthetized by IP injections of avertin. Next, we exposed/cleaned the left carotid artery, before measuring its baseline blood flow using a 0.5-mm micro-flow probe (Transonic Systems Inc). After the blood flow stabilized, 7.5% ferric chloride (FeCl₃) was applied onto a 1-mm diameter filter paper disc. The filter paper was placed immediately onto the left carotid artery for a total of 3 minutes. The carotid artery blood flow was monitored until its stoppage ([stable occlusion] ie, zero blood flow for a total of 2 minutes), with the monitoring lasting no longer than 30 minutes. The findings were documented, with the time to the carotid artery blockade (occlusion) calculated as the difference in time between stable occlusion and removal of the FeCl₃ filter paper. As indicated, a 30-minutes occlusion time served as the cutoff time for statistical analysis purposes.