Fitc dextran4d

As described previously¹⁶ , one day after TBI, mice were gavaged with 0.4 ml of 22mg/ml FITC-dextran4D (Sigma-Aldrich, St. Louis, MO) in phosphate buffer saline. Three hours post gavage with FITC-dextran, mice were sacrificed. Entire luminal contents of stomach, the upper small intestinal section #1, the middle small intestine section #2, the distal small intestine section #3, and large intestine were collected. Luminal contents were weighed and suspended in PBS (weight mg X 5uL PBS ) and sonicated (XL-2000 Misonix) until the solution was homogenous. Homogenates were centrifuged at 8000rpm for 10min at 4°C, supernatants were collected, and read spectrophotometrically at 480nm excitation and 520nm emission wavelengths to determine FITC-dextran concentration and were used for analysis of intestinal transit.

Male C57/BL6 mice (22-25 g) were obtained from Charles River Laboratories (Wilmington, MA). Rag1−/− mice and wildtype mice were obtained from Jackson laboratories (Bar Harbor, ME). IL-6 enzyme-linked immunosorbent assay (ELISA) kit was obtained from BD Biosciences (San Diego, CA). IL-17 and KC ELISA kits were obtained from R&D Systems (Minneapolis, MN). IL-22 and IL-18 ELISA kits, PE IL-22 (Clone 1H8PWSR, Catalog # 12-7221-82, eBioscience™), eFluor450 IL-17 (Clone eBio17B7 Catalog # 48-7177-82 eBioscience™) and APC-eFluor780 CD3 antibodies (Clone 17A2Catalog # 47-0032-82, eBioscience™), and Fixable Viability Dye eFluor506 were obtained from eBioscience (San Diego, CA). Cell Death Detection ELISA kit was obtained from Roche (Catalog # 11920685001, Indianapolis, IN). FITC-dextran (4D) was obtained from Sigma-Aldrich (St. Louis, MO). Primers to CYP1A1 and β-actin, MirVana miRNA Isolation Kit, High Capacity cDNA Reverse Transcription Kit, and TaqMan Gene Expression Master Mix were obtained from Life Technologies (Grand Island, NY). AHR agonist, 6-formylindolo (3, 2-b) carbazole (FICZ) was obtained from Abeam (Cambridge, MA).