Conventional IL-10−/− mice (C57BL/6j background) were reared in the Forschungsinstitute für Experimentelle Medizin, Charité—University Medicine Berlin (Berlin, Germany), housed in cages equipped with filter tops under standard conditions (22–24 °C room temperature, 55 ± 15% humidity, 12 h light/12 h dark cycle) in an experimental semi-barrier facility and had free access to autoclaved standard food pellets (ssniff R/M-H, V1534-300, Sniff, Soest, Germany) and tap water. Female and male 3-week-old litter-mate mice were treated with broad-spectrum antibiotics, in order to deplete the commensal gut microbiota as reported earlier [41 (link),54 (link)]. In brief, mice were transferred to sterile cages (maximum of 4 animals per cage) and treated for eight weeks with an antibiotic cocktail consisting of ampicillin plus sulbactam (1 g/L; Dr. Friedrich Eberth Arzneimittel, Ursensollen, Germany), imipenem (250 mg/L; Fresenius Kabi, Bad Homburg, Germany), vancomycin (500 mg/L; Hikma Pharmaceuticals, London, UK), metronidazole (1 g/L; B. Braun, Melsungen, Germany), and ciprofloxacin (200 mg/L; Fresenius Kabi, Bad Homburg, Germany) added to autoclaved drinking water (ad libitum). Microbiota-depleted mice were continuously kept and handled under strict aseptic conditions. Three days before infection, the antibiotic treatment was withdrawn, and mice received autoclaved tap water instead.
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