2.10. Middle Cerebral Artery Occlusion (MCAO) Surgery

Transient ischemic stroke was induced by temporarily blocking the origin of middle cerebral artery as described in previous studies [6,22]. Briefly to induce transient ischemia, a ventral, midline incision was made under isoflurane anesthesia. Thereafter, the right external carotid artery was incised before inserting a 6.0 mm silicone-coated nylon filament (Doccol Corporation, Sharon, MA, USA) from the internal carotid artery bifurcation via the external carotid artery stump. Rectal temperatures were maintained at ~37 °C with the help of heating pads during the procedure. We confirm successful occlusion by laser doppler, reflecting <15% residual middle cerebral artery flow with the return of flow to 85% of baseline upon reperfusion. After an hour of occlusion, mice were perfused for predetermined time of 48 or 72 h.