Journal: The World Allergy Organization Journal

Article Title: Enhanced allergic Eustachian tube response in surfactant protein D knockout mice induced by ovalbumin

doi: 10.1016/j.waojou.2025.101061

Figure Lengend Snippet: Immunostaining of mucins Muc5ac and Muc5b in different groups of mice treated with OVA or PBS. (A) Immunohistochemical staining showing the secretion of Muc5ac by goblet cells in the midsection of the ET across 4 groups of mice (n = 4). A prominent mucin plug is observed in the lumen of the SPD-KO-OVA group (arrow). Scale bar: 100 μm, 200X. (B–C) The results of immunostaining quantitative analysis show a positive correlation between Muc5ac and Muc5b protein expression levels and the integrated optical density values in specific regions. The SPD-KO-OVA group exhibits significantly higher expression of Muc5ac and Muc5b compared to the WT-OVA group (two-way ANOVA, n = 4). (D) Immunofluorescence staining showing the secretion of Muc5b by goblet cells and glands in the midsection of the ET across 4 groups of mice (n = 4). Pink dashed areas indicate goblet cells of the ET, and white dashed areas represent glandular regions. DAPI (blue), Muc5b (green). Scale bar: 100 μm. G: glands. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001 (intergroup comparisons)

Article Snippet: Sections were blocked at room temperature with goat serum for 45 min. After blocking, rabbit anti-Muc5ac antibody (CST, #61193, 1:500) was applied, and sections were incubated at 4 °C for 14 h. The following day, sections were warmed to room temperature for 30 min, washed 3 times with TBS, and processed according to the instructions of a commercial secondary antibody kit (Beijing Zhongshan Golden Bridge Biotechnology, Cat# PV-6000D).

Techniques: Immunostaining, Immunohistochemical staining, Staining, Expressing, Immunofluorescence

Journal: The World Allergy Organization Journal

Article Title: Enhanced allergic Eustachian tube response in surfactant protein D knockout mice induced by ovalbumin

doi: 10.1016/j.waojou.2025.101061

Figure Lengend Snippet: Effects of exogenous SPD protein treatment on morphological and functional outcomes in SPD-KO-OVA mice. (A) Representative H&E-stained cross-sectional views of the mid-portion of the ET in PBS control and rmSPD treatment groups (n = 4). Circles indicate eosinophil infiltration. Scale bar: 100 μm. (B) Immunofluorescence staining of goblet cells and glands secreting Muc5b in the ET of both groups (n = 4). Pink dashed outlines mark goblet cells; white dashed outlines denote glands. Nuclei (DAPI, blue), Muc5b (green). (C) Immunohistochemical staining of Muc5ac secretion by goblet cells in the ET (n = 4). Scale bar: 100 μm. (D) Serum total IgE levels measured by ELISA (n = 8). (E) Comparison of the ACNP function in the ET between groups. (F–G) Analysis of average epithelial thickness and eosinophil counts across 3 experimental groups (n = 4). (H–I) Semi-quantitative analysis of immunostaining intensity showing significantly lower expression of Muc5ac and Muc5b in the rmSPD treatment group versus controls (n = 4). (J) ACNP rates were significantly higher in the rmSPD treatment group compared to untreated and control groups (n = 4). rmSPD: recombinant mouse SPD protein. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001 (intergroup comparisons)

Article Snippet: Sections were blocked at room temperature with goat serum for 45 min. After blocking, rabbit anti-Muc5ac antibody (CST, #61193, 1:500) was applied, and sections were incubated at 4 °C for 14 h. The following day, sections were warmed to room temperature for 30 min, washed 3 times with TBS, and processed according to the instructions of a commercial secondary antibody kit (Beijing Zhongshan Golden Bridge Biotechnology, Cat# PV-6000D).

Techniques: Functional Assay, Staining, Control, Immunofluorescence, Immunohistochemical staining, Enzyme-linked Immunosorbent Assay, Comparison, Immunostaining, Expressing, Recombinant