Murine experimental stroke models

Surgical anesthesia was induced with 4-5% isoflurane and maintained with 1-2% isoflurane via face-mask in oxygen-enriched air. We performed focal cerebral ischemia by transient middle cerebral artery occlusion (tMCAO) or permanent middle cerebral artery occlusion (pMCAO) with a 6.0 monofilament suture (Doccol, Sharon, Massachusetts). We used laser Doppler flowmetry (Moor instruments, United Kingdom) to detect regional cerebral blood flow and confirm a successful occlusion (>70% decrease in flow). Rectal body temperature was maintained at 37 ± 0.5 °C during surgery. Randomized controls were used under a common protocol that collectively assessed 353 mice, including 42 mice with sham surgery, 40 mice with pMCAO, and 271 mice with tMCAO. All surgeries were performed by one surgeon who was blinded to pre-treatments. Neurological deficits of 311 stroke mice were recorded at 6-hour, 24-hour, and 48-hour end points. Infarct volume of 162 stroke mice was determined at 24-hour or 48-hour end points using 2,3,5-triphenyl-2H-tetrazolium chloride (TTC; Sigma) staining.