Cerebral ischemia was induced by 90 min of reversible MCAO under isoflurane anesthesia, as described previously [26 (link), 32 (link), 56 (link)]. In brief, a midline ventral neck incision was made, and unilateral right MCAO was performed by advancing a 6.0 silicone-coated nylon monofilament (Doccol Corporation, CA) into the internal carotid artery 6 mm from the internal carotid–pterygopalatine artery bifurcation via an external carotid artery stump. Rectal temperatures were monitored with a temperature control system (Fine science tools, Canada) and temperature was maintained with an automatic heating pad at ~37 °C during surgery and ischemia. Cerebral blood flow measurements by laser Doppler flowmetry (DRT 4/Moor Instruments Ltd, Devon, UK) confirmed ischemic occlusion (reduction to 85 % of baseline) during MCAO and restoration of blood flow during reperfusion. Surgical controls are used for molecular analysis, a sham surgery in which the suture was not advanced into the internal carotid artery (controls). All mice are allowed to emerge from anesthesia after the initial suture advancement into the MCA and placed back into their home cage until re-anesthetized for reperfusion. This allows for the assessment of intra-ischemic behavioral deficits to confirm successful suture placement.