Transient Middle Cerebral Artery Occlusion (tMCAO) and Drug Treatment

The healthy male SD rats were randomly divided into a series of groups for transient middle cerebral artery occlusion (tMCAO) (n = 12–15 for each group of successfully treated rats). Firstly, the rats were treated with anesthesia in an isoflurane chamber with 3.5% isoflurane, and then 2% isoflurane was maintained through a mask in the operation. During the surgery, the animals were placed on a heating device to ensure normal body temperature (37°C). After accurate separation of the right common carotid artery (CCA), internal carotid artery (ICA), and external carotid artery (ECA), a monofilament nylon suture (about 0.24 mm in diameter) with a rounded tip was inserted through the ECA stump into the ICA and gently advanced to the MCA. Then, in order to monitor blood block, the Laser Speckle Imaging system (moorFLPI-2^™) was used to detect whether the baseline of brain blood flow was >75% reduced (Figure S2). Two hours after cerebral ischemia, filament was removed to restore blood flow (reperfusion). All rats had free access to food and water. At 3 h and 24 h after the ischemia, the separated groups of rats were intravenously injected with indicated dose of CBL (dissolved in saline) or saline alone. In total, 120 rats went through tMCAO operation, among which 6 rats were excluded for hemorrhagic transformation, 10 for unsuccessful occlusion and 9 rats died during the operation.