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Quantification of cytokines by ELISA
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Approximately 100 mg of colon tissue was harvested and homogenized in PBS containing protease inhibitors (Thermo Fisher Scientific). Samples were centrifuged for 10 min at 2000 × g, and supernatants were used for quantifying TNF-α, IL-6, IL-1β, IL-10, KC (Cxcl1), and MIP-2 (Cxcl2) using the Duo Set ELISA kit (R&D Systems; Minneapolis, MN, USA). Data were normalized to the total protein concentration determined using the Bradford protein assay.
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