Mice in the three diet groups were randomly divided into three additional groups (n=8 per group): controls, OVA and OVA + ozone. The mice in the OVA and OVA + ozone groups were sensitized intraperitoneally (IP) with 20 µg OVA (Grade V, Sigma-Aldrich) diluted in 0.2 mL of Dulbecco’s phosphate buffered saline (PBS) and 2 mg aluminum hydroxide (Sigma-Aldrich) in a total volume of 20 mL on days 0, 7, and 14, and challenged via aerosol nebulization with 1% OVA for 30 min each day from day 21 to day 25. Mice in the control group were treated with PBS at both timepoints in the same manner as previously described (33 (link)). After the OVA challenge, the same number of mice in the OVA + ozone group were exposed to 2.5 ppm ozone for 2 h daily from day 21 to day 25 as previously described (34 (link), 35 (link)). The control mice were exposed to room air during this period.
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