Sk38 2ml tubes

Tissue samples were weighed and were homogenized in 500 μl PBS, after which lysis was completed by addition of lysis buffer (20 mM Tris HCl (pH 7.4), 200 mM NaCl, 1% Nonidet P-40) and incubation for 10 minutes on ice. Full speed centrifugation for 30 minutes cleared the homogenate and supernatant was used for further analysis. Mouse cytokines in cell culture supernatants and tissue homogenates were determined by magnetic bead-based multiplex assay using Luminex technology (Bio-Rad) and type I IFN levels were analyzed using eBioscience IFNα/IFNβ Procartaplex, all according to the manufacturer’s protocols. In S5E Fig, IL-1β was detected by the BD OptEIA ELISA kit (BD biosciences) according to the manufacturer’s protocol. Cytokines from tissue homogenates were normalized to weight of tissue, while cytokines from cell culture supernatants were expressed as concentration per ml of cell culture medium. For measuring Lipocalin-2 levels, fecal pellets were weighed and were homogenized in 500μl PBS using sterile soil grinding SK38 2ml tubes (Bertin Technologies). Stool homogenates were collected in 1.5 ml eppendorf tubes and cleared upon full speed centrifugation for 30 minutes. Lipocalin-2 levels in stool supernatants were then analyzed using the mouse Lipocalin-2/NGAL duoset ELISA (R&D systems) according to the manufacturer’s instructions, and were normalized per mg of stool.