Studies were conducted with approval of the University of Colorado Denver and Johns Hopkins University Institutional Animal Care and Use Committees. Housing rooms are maintained at 22 °C, 30–40% humidity, and a 14/10 (h/h) light/dark cycle and at least 12 fresh-air changes per h. Male and female BALB/cJ wild type mice were purchased from the Jackson Labs (Bar Harbor, ME). Muc5ac−/− mice were previously crossed onto a congenic BALB/cJ strain background15 (link). Animals were housed under specific pathogen-free conditions and used in allergic asthma studies beginning at ages 6–8 weeks.
To induce allergic inflammation and mucin overproduction, mice were challenged using aerosolized Aspergillus oryzae extract15 (link) (AOE; Sigma), as shown in Supplementary Fig. 2. Mice received four weekly challenges, and endpoint analyses were studied 48 h after the last AOE challenge. To induce mucolysis, mice and mucus samples were exposed to tris(2-carboxyethyl)phosphine (TCEP, neutral pH solution, Thermo Scientific, Cat. no. 77720). Mice exposed to saline challenges were used as controls.
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