A total of one hundred and thirty heathy male C57BL/6 mice (20-25 g) were purchased from the Beijing HFK Bioscience Corporation, China. Mice were maintained 5 per cage in a temperature-controlled room (22 ± 2°C), at a 12 h light-dark cycle and a relative humidity of 60 ± 10%. The mice were randomly allocated to 3 groups: sham, MCAO, and MCAO+RIPC group. Then, all mice were euthanized at 24 h or 72 h post-MCAO, respectively. In order to assess neurological deficit, infarct size, and ELISA analyses,n = 6 for each group were utilized. For real-time PCR, n = 4 per group. For immunofluorescence, flow cytometry analysis, and Western blot, n = 3 for each group. Twenty-two mice died from intracranial hemorrhage, and the data from these mice were excluded in the analysis. The protocols for all animal experiments were approved by the Institutional Animal Care and Use Committee of China Medical University (Shenyang, China).

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