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<p>Mice We used females from 41 BXD RI strains and both parental strains—B6 and D2. Mice were between 8 and 12 weeks of age when infected. They were housed and maintained on a 12:12 light/dark cycle, with ad libitum access to food and water. Virus Original stocks of mouse-adapted A/Puerto Rico/8/34 (H1N1, PR8M) virus were obtained from Stefan Ludwig, University of Münster (28). Virus stocks were propagated in the chorioallantoic cavity of 10-day-old pathogen-free embryonated chicken eggs for 48 h at 37°C as described previously (29). Viral titer was determined using a focus-forming unit (FFU) assay as described previously (29). Infection of Mice Animals were anesthetized by intraperitoneal injection of ketamine/xylazine (10 % (v/v) of 100 mg/ml ketamine and 5 % (v/v) of 20 mg/ml xylazine in 0.9 % (w/v) NaCl with a dose adjusted to body weight (200 μl/20 g body weight). Infection was performed by intranasal application of virus solution in 20 μl sterile phosphate-buffered saline (PBS), with a PR8M dosage of 2×103 FFU. Mice were bred and infected at the animal facilities at UTHSC</p>
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