Experiments were performed on 6‐ to 8‐week‐old male adult Sprague‐Dawley (SD) rats (220‐300g), as supplied by the laboratory animal centre of Shihezi University. Then, rats were divided into 9 groups randomly (n = 15‐20 each group): (1) Sham group, (2) I/R group, (3) shMEF2D + IR group , (4) shScr + IR group , (5)IR + ISO group, (6) shMEF2D + IR+ISO group, (7)shScr + IR+ISO group , (8) XMD8‐92 + IR+ISO group (X + IR+ISO) and (9) DMSO + IR+ISO group (D + IR+ISO). XMD8‐92 (5 μg/kg) was injected into the right ventricle at 30 min before MCAO (coordinate: A‐P −1mm, M‐L 1.5 mm, D‐V −4.5 mm).

We used a total of 187 rats. Rats with massive haemorrhage or complications of the middle cerebral artery during surgery were excluded. In all of the groups, the mortality rate was 6.4% (12 of 187), and the exclusion rate was 4.4% (7 of 158). The processing procedures of all animals were according to the National Institutes of Health (NIH) Guide for the Care and Use of Experimental Animals (80‐23, 1996). Study design and experimental methods were conformed to the Animal Ethics Committee of the First Affiliated Hospital of Shihezi Medical College.

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