Prepared sections of nasal tissue were deparaffinized and antigen was retrieved by incubation in citrate buffer (pH 7.4) under high heat and pressure. Endogenous peroxidase activity was blocked by incubation of sections with 3% hydrogen peroxide in phosphate buffer solution (PBS) and non-specific staining was blocked with normal goat serum for 40 min at room temperature. The tissue sections were incubated overnight at 4°C in a humidified chamber with anti-T-bet (1:1600) or anti-GATA3 (1:500) antibodies diluted in PBS. After washing, the sections were incubated with biotin-conjugated goat anti-rabbit or anti-mouse IgG secondary antibodies for 30 min at room temperature. Antibody binding was visualized using a 3,3′-diaminobenzidine detection system. Slides were digitized using a Panoramic SCAN digital slide scanner with a Zeiss plan-apochromat objective (magnification 20×, aperture 0.8) and a Hitachi (HV-F22CL) 3CCD progressive scan color camera (resolution 0.2325 μm/pixel).

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