Journal: Microbiome
Article Title: Role of nasal microbiota in regulating host anti-influenza immunity in dogs
doi: 10.1186/s40168-025-02031-y
Figure Lengend Snippet: Dysbiosis of nasal microbiome diminishes the antiviral response within the nasal cavity. A Volcano plot of the differentially expressed genes (DEGs) between the Nor, WT, and Abx groups in the nasal cavity at 8 dpi with the influenza virus (T3). B The Venn diagram illustrates the shared and unique DEGs among the Nor, WT, and Abx groups in the nasal cavity associated with influenza virus infection at 8 dpi. C Clustering analysis performed on the DEGs in nasal tissue from the Nor, WT, and Abx groups using the Fuzzy-c means (FCM) algorithm. D The k -means clustering analysis of the DEGs using the STING online database ( https://string-db.org/ , Accessed 20 May 2023) with a minimum required interaction score of 0.4. The DEGs in the nasal tissue between the WT and Abx groups were primarily categorized into five clusters. These clusters predominantly encompass the genes associated with IFN-mediated antiviral responses (Cluster 1), mucin generation (Cluster 2), barrier formation (Cluster 3), inflammation (Cluster 4), and autophagy pathway (Clusters5). E The heatmap illustrates the DEGs among five clusters between the WT and Abx groups. All genes were normalized based on the corresponding genes in the Nor group before conducting differential analysis. All corresponding significance levels are determined by the DESeq2 package (Benjamini-Hochberg). F The mRNA transcript levels of cytokines (IFNβ1, ISG15, Mx1, OAS1, IL1, IL6, TNFα, and Caspase3) in the blood with influenza virus at 8 dpi were measured in the Nor, WT, and Abx groups (one-way ANOVA test, Tukey’s HSD). G Representative images of immunofluorescence staining of nasal tissue show ZO-1/TJP1 protein (red) and cell nuclei (blue, DAPI). ZO-1/TJP1 exhibited uniform expression in the Nor group, but presented heightened expression at the injury site in the WT group (indicated by an arrow). In contrast, the Abx group displayed an overall diminished expression of ZO-1/TJP1 protein at the injury site
Article Snippet: The sections were sequentially incubated with the primary antibody at 4 °C overnight and anti-rabbit secondary antibody conjugated with Alexa Fluor 488 (Proteintech, China) or anti-mouse secondary antibody conjugated with Alexa Fluor 555 (Proteintech, China) at room temperature for 1 h. In this study, the primary antibodies targeting the following proteins were used at 1:100 dilution: influenza virus nucleoprotein (NP; laboratory preparation), tight junction protein 1 (TJP1/ZO-1; Bioss, China), microtubule-associated protein 1 light chain 3 beta (MAP1LC3B/LC3B; Abmart, China), or sequestosome 1 (SQSTM1/p62; Abmart, China).
Techniques: Virus, Infection, Immunofluorescence, Staining, Expressing
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