Tissue Cytokine Measurements

Following DSS or water administration, flash-frozen whole colon was homogenized in Tris lysis buffer containing a protease and phosphatase inhibitor cocktail (Thermo Fisher), using a handheld tissue homogenizer. The concentration of cytokines in tissue lysates was measured using the V-Plex Pro-Inflammatory Panel Mouse kit (Meso Scale Discovery, Rockville, MD) per the manufacturer’s instructions. Signal detection was performed using a Sector Imager 2400 (Meso Scale Discovery). A Bicinchoninic Acid (BCA) Protein Assay Kit (Thermo Fisher) was used to determine the amount of protein loaded for each sample. Cytokine concentrations in tissue homogenate were normalized to total protein.