2.3. Transient middle cerebral artery occlusion (tMCAO)

Mice were anesthetized with isoflurane (induction: 5%; maintenance: 1.5–2%) in oxygen (0.05 L/min) and medical air (1 L/min). After sterile preparation and draping, tMCAO was performed as previously described [1] with minor modifications. Briefly, the right common carotid artery (CCA) was exposed, and a 7-0 nylon suture tipped with dental adhesive was introduced into the CCA until laser Doppler signal (Periflux System 5000, Perimed, Inc., Ardmore, PA) from a probe positioned over the MCA territory dropped by ≥80%. During MCAO, rectal temperature was maintained between 36.5 and 37.5 °C with a homeothermic system (Harvard Apparatus, Holliston, MA). After 30 min, the occluding thread was removed, and the neck was sutured. For the rat stroke model, young adult male Wistar rats underwent MCAO as previously described [2].