*4.3. Global Cerebral Ischemia Surgery*

Male C57BL/6J mice (aged 2–3 months, weight 20–25 g) from DBL (Chungcheongbuk-do, Eumseong-gun, Korea) were used as controls. The mice were anesthetized with 2% isoflurane in a 75:25 mixture of oxygen and nitrous oxide. Core temperature was maintained at 36.7–37.5 ◦C with a homeothermic blanket control unit (Harvard Apparatus, Holliston, MA, USA). Bilateral common carotid arteries (BCCAs) were exposed through a midline neck incision. The BCCAs were loosely encircled with a 4/O silk suture before the occlusion. Aneurysmal clips were used to occlude BCCAs. Mice were subjected to common carotid artery occlusion for 30 min while anesthetized with 1% isoflurane [71–73]. The aneurysmal clips were removed, and the BCCAs were inspected for normal recovery of blood flow after the end of the 30 min ischemic period. Anesthetics were discontinued following the suture of the skin incision. When mice confirmed spontaneous respiration, they were returned to a recovery room retained at 37 ◦C. Sham-operated animals received the same neck skin incision under isoflurane anesthesia without BCCA occlusion.
