Cytokine-cytokine receptor interactions in the highly pathogenic avian influenza H5N1 virus-infected lungs of genetically disparate Ri chicken lines

Figure 1. Heat map of the individual H5N1-infected samples from the lung tissue of resistant and susceptible Ri chickens at three days post-infection. A green color indicates DEGs that have higher expression levels in resistant chickens while a red color indicates DEGs that have lower expression levels in resistant chickens, as calculated from the expression values in log10 (FPKM) units. H5N1, influenza A virus subtype H5N1; DEGs, differentially expressed genes; FPKM, fragments per kilobase of exon model per million reads mapped.

Figure 2. Gene ontology (GO) functional analysis. The enriched biological terms include: (A) Top 10 SEA: GO Biological process, (B) Top 10 SEA: GO Cellular component, and (C) Top 10 SEA: GO Molecular function from the 972 DEG in Ri chicken lines obtained using the criteria (|fold-change| ≥2) ∩ (p≤0.05). GO, gene ontology; SEA, singular enrichment analysis terms; DEG, differentially expressed genes.

Figure 3. Six Kyoto encyclopedia of genes and genomes functional pathways of differentially expressed genes in Ri chickens of the selected differentially expressed genes.

Figure 4. Location of differentially expressed genes associated with cytokine-cytokine receptor interactions in the lungs of resistant H5N1-infected Ri chickens compared with susceptible chickens. The location of 33 significantly different expressed genes were marked. A green color indicates DEGs that have higher expression levels in resistant chickens while a red color indicates DEGs that have lower expression levels in resistant chickens (CXCL14). H5N1, influenza A virus subtype H5N1; DEG, differentially expressed genes.

Figure 5. Interactions of the 33 differentially expressed genes in the lungs of Ri chicken lines. This interaction analysis was conducted using the Pathway Studio program.

Figure 6. Analysis of the expression of genes associated with cytokine-cytokine receptor interactions in resistant H5N1-infected chickens compared with susceptible H5N1-infected chickens. The expression patterns of these genes were compared between real-time qPCR and RNA sequencing data. Relative quantitation data of qRT-PCR are represented as mean±SEM normalized to GAPDH using the 2−ΔΔCt method. H5N1, influenza A virus subtype H5N1; qRT-PCR, quantitative real-time polymerase chain reaction; SEM, standard error of the mean; GAPDH, glyceraldehyde-3-phosphate dehydrogenase. Data are expressed as mean±SEM of three independent experiments: * p<0.05, ** p<0.01, and *** p<0.001.