Figure 3. IAV profoundly alters the airway epithelium.

HBECs were infected with 100,000 PFU of IAV for 72 hours. (A) HBECs were stained for cilia (white), F-actin (green), or IAV (red) and mounted in DAPI Fluoromount (blue). Images were taken at ×40 magnification on a Nikon A1R-HD25. Scale bar: 20 μm. Image is representative of 3 replicates. (B) Principal component analysis (PCA) of the transcriptome of HBECs with and without IAV. (C) Cnetplot of enriched gene ontology (GO) terms after infection with IAV, with the size of each dot representing the number of differentially expressed (DE) genes in the category and the color indicating log₂(fold change). The cnetplot was generated using the R package ClusterProfiler v4.0 (D–F) Heatmap of ion channel (GO: 0015276), mucociliary clearance (GO: 0120197), complement activation (GO: 0006956), and response to a molecule of bacterial origin (GO: 0002237) gene expression in HBECs with and without IAV infection. Transcriptomic analysis was conducted on 3 replicates. Green indicates uninfected and orange indicates IAV infected. DE genes were calculated with a false discovery rate of ≤0.05 and an absolute log₂(fold change) of ≥1. Plots in D–G are composed of DE genes from the respective category.