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.
To induce allergic inflammation and mucin overproduction, mice were challenged using aerosolized Aspergillus oryzae extract15 (link) (AOE; Sigma), as shown in Supplementary Fig.