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Immediately prior to sacrifice, blood was collected via retro-orbital sinus with heparinized capillary tubes, placed on ice, and centrifuged (10,000g for 10 min at 4°C) (46). The supernatant was removed, and plasma IL-6 concentrations were determined using an enzyme-linked immunosorbent assay kit according to the manufacturer’s instructions (Catalog #KMC0061, ThermoFisher Scientific, Waltham, MA).
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