Nissl staining was used to measure volume of infarct [26]. Briefly, the coronal cryosections of the brain (20-μm thick, 1-mm intervals) were stained with cresyl violet (Cat. G1430; Solarbio, Beijing, China) for 10 min at 60 °C. Images were captured and analyzed using National Institute of Health (NIH) ImageJ software (Version 1.46r; Bethesda, MD, USA). The infarct area was calculated as non-infarcted contralateral hemisphere area minus surviving area of infarcted ipsilateral hemisphere. Infarct (V1) and contralateral hemisphere (V2) volumes were computed by numeric integration of sequential regions, and the results were presented as: Infarct volume (%) = V1/V2 × 100%.
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