Ne uo7

Female BALB/c mice (n = 30 in each group), 6 weeks of age and free of mouse-specific pathogens, were obtained from Orient Bio (Orient Bio Inc., Seongnam, Republic of Korea). The particle inhalation methods for mice were conducted as in previously studies [7 (link),32 (link)]. The mice were housed throughout the experiments in a laminar flow cabinet and maintained on standard laboratory chow ad libitum. The mice were exposed to 100 μg/m³ DEPs in a closed-system chamber attached to an ultrasonic nebulizer (NE-UO7; Omron Corporation, Tokyo, Japan) with an output of 1 mL/min and a 1 to 5 µm particle size. The control mice were administered and exposed to a saline solution alone. The mice were exposed to DEPs for 1 h a day for 5 days a week from 4 and 8 weeks, and on days 27 and 55 AHR was measured. On days 28 and 56, BALF was collected and tissue was processed for protein immunoblotting. After ligation of the right main bronchus, the left lung was fixed with 4% paraformaldehyde in phosphate-buffered saline and paraffin-embedded for immunohistochemistry. All experimental animals in this study were used according to the guidelines of the Institutional Animal Care and Use Committee (IACUC) at Soonchunhyang University Medical School.

All animal experiments were conducted according to the guidelines approved by the Institutional Animal Care and Use Committee at Soonchunhyang University Medical School (SCHBC-Animal-2014-013). All experimental protocols were approved by the Institutional Animal Care and Use Committee of Soonchunhyang University Medical School. Specific pathogen-free, 6-week-old female BALB/c mice were used for animal experiments. DEPs (National Institute of Standards and Technology, Gaithersburg, MD, USA) were sterilized by autoclaving and were suspended in saline solution for 30 min before exposure to mice. The DEP-treatment group (n = 8) inhaled 100 μg/m³ DEPs via an ultrasonic nebulizer (NE-UO7; Omron Corporation, Tokyo, Japan) for one hour per day, five days a week, for four weeks, with an output of 1 mL/min and 1- to 5-µm particle size^{42 (link)}. The control group (n = 8) was treated with saline solution under the same conditions as the experimental group. After DEP exposure, animals were sacrificed to gain access to the nasal cavity including the nasal septum and turbinate. The head was separated under anesthesia. After the jaw and nose were removed, nasal tissues including the nasal septum and turbinate between the incisive papilla and second molar were collected.