Fig. 3

Top 30 differentially expressed genes. Increased tidal volume leads to increased differential expression. Shown are the top 30 genes with the highest beta value from RNA sequencing of mouse lung tissues. Up regulated genes showed increased expression with increased tidal volume, while down regulated genes decreased in expression with increased tidal volume (A). Genes involved in glutathione production were significantly upregulated. The expression was shown to be tidal volume dependent (B). Real time PCR was done using lung tissue from ventilated mice and compared to SB controls (C). The top three genes from sequencing analysis were confirmed to be expressed in a tidal volume dependent manner. Immunoblotting from SB control, 10, 20 and 30 mL/kg tidal volume ventilated mice was performed for Keap1 and Actin. Actin used as a loading control and quantification carried out using FIJI (D). Significant difference from the SB controls are shown (P ≤ 0.05 = *; P ≤ 0.01 = **; P ≤ 0.001 = ***)