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For electron microscopy examination, liver tissues were prefixed after harvesting immediately with 1.5% glutaraldehyde and 0.8% paraformaldehyde (0.1 mol/L cacodylate buffer) at room temperature and postfixed in an aqueous solution of 1% OsO 4 and 1.5% K 4 (FeCN) 6

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Primary antibodies for Occludin and ZO-1 were purchased from Abcam (Cambridge, United Kingdom)