Nutra gel

The experiments outlined were performed on B6 (C57Bl/6, The Jackson Laboratory, Bar Harbor, ME) mice. Mice were housed in a pathogen-free facility, caged individually, had access to a 24% protein mush-based feed, Nutragel (Bio-Serv, Frenchtown, NJ), and autoclaved reverse osmosis water. Mice were kept in a temperature (22 ± 2˚C) and humidity (30–70%) controlled environment with a 12-hour light cycle. Mouse adapted human influenza virus A/PR/8/34 (PR8) for infection was produced as described previously [14 (link)]. Eight-week-old mice were exposed to an infective dose of PR8 (500 TCID₅₀) in an aerosolization chamber (Glas-Col, Terre Haute, IN) [15 ]. Uninfected control mice were matched for food intake. Mice were sacrificed on Day 5 of infection by 5% Isoflurane inhalation with cervical dislocation. All tissues were isolated and stored at −80˚C until use. Single dose liposomal clodronate (200 μL/animal i.p., Encapsula NanoSciences LLC, Brentwood, TN) was used to deplete Kupffer cells. All animal care and procedures were carried out according to the criteria outlined in the “Guide for the Care and Use of Laboratory Animals” prepared by the National Academy of Sciences and published by the National Institutes of Health (NIH publication 86–23 revised 1985) and were authorized by the Animal Care and Use Committees of the National Human Genome Research Institute.

All animal procedures were approved by the University of Alabama at Birmingham Institutional Animal Care and Use Committee. Systemic, constitutive KL–deficient (KO, 129S1/SvImJ) and systemic, constitutive KL–overexpressing (OE, C57BL/6J) mice were obtained from M. Kuro–o (University of Texas Southwestern). We purchased Thy1–GFP (C57BL/6J) from the Jackson Laboratory (Stock #007788; Bar Harbor, ME). All mice were allowed access to food and water ad libitum and housed at 26.6°C with humidity maintained at ≥40%. Beginning at 5-weeks of age, we supplemented KL–deficient mice with Bacon Softies or Nutra–Gel (BioServ, Frenchtown, NJ). We previously reported no effect of sex on KL-deficient mouse behavioral performance (Laszczyk et al. 2017 (link)). Herein, sex balanced groups of animals were used where possible, but sex was not determined in cultured neurons taken from P1 pups.

KL-deficient (KO,129S1/SvImJ) and overexpressing (OE,C57BL/6J) lines of mice are from M. Kuro-o (University of Texas Southwestern and Jichi Medical University, Japan). Strain-specific, wild-type (WT) controls were used for all experiments. All mice were housed with free access to food and water at 26.6°C, humidity maintained above 40%. KO mice were supplemented with Bacon Softies or Nutra-gel (BioServ, Frenchtown, NJ) beginning week 5. All procedures were approved by the University of Alabama at Birmingham Institutional Animal Care and Use Committee.

Mice were placed into individual metabolic cages, and a gel diet containing nutrients, water and sodium (Nutra-Gel, Bio-serv, Frenchtown, NJ) was provided. Sodium ingestion and excretion were quantitated daily to allow quantitation of sodium balances.