*2*.*2*. *MCAO/Re*

The experimental MCAO/Re rat model was prepared as described previously [25]. The rats were anesthetized with halothane (4% for induction and 1.5% for maintenance) under spontaneous respiration. After a midline incision on the neck, the right common carotid artery was isolated under an operating microscope. All branches of the external carotid artery were ligated. The tip of a 4–0 surgical nylon monofilament rounded by flame heating was inserted through the internal carotid artery and advanced to occlude the origin of MCA. The rectal temperature was maintained at 37 °C with a heat lamp and a heating pad during the operation. After 2 h of occlusion, the filament was withdrawn to enable reperfusion. The distance from bifurcation of the common carotid artery to the tip of the suture was approximately 20 mm in all rats. Cerebral blood flow was detected using a Laser Doppler flowmetry (ATBF-LC1, Unique Medical, Tokyo, Japan), and approximately 50% reduction of its baseline associated with MCAO was ascertained in the rats. Then, the rats were allowed to recover from anesthesia at room temperature and were killed after 24 h of reperfusion. Sham operation involved the same manipulations but insertion of the filament.
