Precellys tissue homogenizer tubes

Intestinal tissue was weighed and homogenized (100 mg tissue/mL) in cell lysis buffer (Cell Signaling Technology, Beverly, MA) with protease inhibitors (Roche) using Precellys tissue homogenizer tubes (Bertin Technologies, Montigny, France). For cytokine bead array, the BD Cytometric Bead Array Mouse Cytokine Kit (560485; BD Biosciences) was used according to the manufacturer’s protocol. Concentrations in tissue lysates were corrected for total protein content as measured by BCA (Pierce, Thermo Fisher Scientific). The concentration of cytokines were determined by sandwich enzyme-linked immunoassay kit (DY460; R&D Systems, Minneapolis, MN).

Intestinal tissue was weighed and homogenized (100 mg tissue/mL) in cell lysis buffer (Cell Signaling Technology, Beverly, MA) with protease inhibitors (Roche) using Precellys tissue homogenizer tubes (Bertin Technologies, Montigny, France). For cytokine bead array, the Cytometric Bead Array Mouse Cytokine Kit (560485; BD Biosciences) was used. Serum amyloid A and myeloperoxidase (MPO) concentration were measured by enzyme-linked immunosorbent assay kits for MPO (HK210-02; Hycult Biotech, Uden, Netherlands) and serum amyloid A (TP 802M; Tridelta Development, Ltd, Maynooth, Ireland). MPO concentrations in tissue lysates were corrected for total protein content as measured by bicinchoninic acid (BCA) assay (Pierce, Thermo Fisher Scientific). The concentration of CXCL12 in the supernatant of fibroblasts was determined using a sandwich enzyme-linked immunosorbent assay kit (DY460; R&D Systems).