Evans blue extravasation assay was performed to assess the permeability of the blood-brain barrier (BBB) after ICH. In brief, a 2% Evans blue (4 ml/kg, Sigma, St. Louis, MO, USA) solution in saline was administered into the mice tail vein 3 h before sacrifice on day 3 after operation. After 3 h circulation, animals were deeply anesthetized and perfused with cold saline through the left ventricle until the colorless fluid outflowed from the right atrium. Afterward, the ipsilateral and contralateral hemispheres were rapidly collected. Then, the hemispheres were weighed and homogenized in 1 ml of 50% ice-cold trichloroacetic acid (Sigma, St. Louis, MO, USA). After centrifugation, the supernatant was collected and mixed with ethanol (1 : 3). A spectrophotometer at a wavelength of 610 nm was used to measure the absorbance. The data were expressed as Evans blue (μg)/tissue (g).

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