NSG mice were obtained from Charles River Laboratories (Sulzfeld, Germany). The mice were kept under specific pathogen-free conditions in individually ventilated cages in a facility controlled according to the Federation of Laboratory Animal Science Association (FELASA) guidelines. The age of the mice was between 6 and 8 weeks.
Following engraftment on day 1, the mice were presensitized by rectal application of 150 µL of 10% ethanol on day 8 using a 1 mm cat catheter as previously described [23 (link)] (Henry Schein, Hamburg, Deutschland). The catheter was lubricated with Xylocain©Gel 2% (AstraZeneca, Wedel, Germany). Rectal application was performed under general anesthesia using 4% isofluran. Post application, the mice were kept at an angle of 30° to avoid ethanol dripping. On day 15, the mice were challenged by rectal application of 150 µL of 50% ethanol following the protocol of day 8. On day 18, the mice were sacrificed. Therapeutic antibodies (150 µL in PBS) were applied i.p. on days 7 and 14, and small-molecule inhibitors (150 µL in PBS or 0.5% methylcellulose gel in PBS (Merck KGaA, Darmstadt, Germany, Firma Cat# M0512) were applied i.p. on days 7–9 and 14–17.
Following engraftment on day 1, the mice were presensitized by rectal application of 150 µL of 10% ethanol on day 8 using a 1 mm cat catheter as previously described [23 (link)] (Henry Schein, Hamburg, Deutschland). The catheter was lubricated with Xylocain©Gel 2% (AstraZeneca, Wedel, Germany). Rectal application was performed under general anesthesia using 4% isofluran. Post application, the mice were kept at an angle of 30° to avoid ethanol dripping. On day 15, the mice were challenged by rectal application of 150 µL of 50% ethanol following the protocol of day 8. On day 18, the mice were sacrificed. Therapeutic antibodies (150 µL in PBS) were applied i.p. on days 7 and 14, and small-molecule inhibitors (150 µL in PBS or 0.5% methylcellulose gel in PBS (Merck KGaA, Darmstadt, Germany, Firma Cat# M0512) were applied i.p. on days 7–9 and 14–17.
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