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For cytokine/chemokine measurements, 17-Plex, duplex and single-plex analyte detection panels were run on the samples using Human Luminex Multiplex Immunoassay kits (R&D Systems)
An equivalent amount of protein from each animal was mixed with 4 sample buffer with or without 2-mercaptoethanol, then separated by SDSpolyacrylamide gel electrophoresis, and transferred electrophoretically to a PVDF membrane (Merck Millipore)
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