Protein lysates were generated from 1-cm proximal colon samples as described previously (Zhou et al., 2023 (link)). PB was obtained by cardiac puncture and allowed to clot for 1-2 hours at room temperature. Following centrifugation at 2000 rpm for 10 minutes (4°C), serum was removed and stored at −80°C until use. Colon lysates and serum samples were subjected to a bead-based multiplex assay (Cytokine & Chemokine 36-Plex Mouse ProcartaPlex Panel 1A, Invitrogen, Waltham, MA) performed on a Luminex 200 platform according to the manufacturer’s instructions. A 96-well plate assay was designed to analyze an array of 36 cytokines and chemokines (CCL2, CCL3, CCL4, CCL5, CCL7, CCL11, CXCL1, CXCL2, CXCL5, CXCL10, G-CSF, GM-CSF, IFN-α, IFN-γ, IL-1α, IL1-β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL12p70, IL-13, IL-15, IL-17A, IL-22, IL-23, IL-27, IL-28, IL-29, IL-31, LIF, M-CSF, and TNF-α). All samples were assayed in triplicate, and standards were measured in duplicate.