Mice were anesthetized and exposed to escalating concentrations of methacholine (0-60 mg/mL, Sigma-Aldrich) via a tracheal cannula, and airway resistance was recorded by using the Buxco Pulmonary Mechanics System (Buxco Electronics, Wilmington, NC). Airway resistance (cm H2O × seconds per mL) at each individual methacholine concentration was recorded and compared with that of PBS after subtracting baseline resistance.
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