Fig. 5

RNASeq analysis comparing WT and ISM1^−/− lungs at 2 months old. Immune-related gene set enriched in ISM1^−/− lungs suggests that ISM1^−/− mice are more prone to allergen-triggered immune responses. (A) Illustration of the RNASeq protocol. Total RNA was isolated from both sides of the lung tissue. (B) Unsupervised hierarchical clustering. (C) Principal component analysis of gene expression shows a distinction between ISM1^−/− samples from the WT. (D) Venn diagram presenting the number of genes that are unique or co-expressed in WT and ISM1^−/−. (E) Validation of selected DE genes by qPCR. (F) Correlation between RNA-Seq and RT-PCR gene expression (R² = 0.96, p < 0.0001). (G) The volcano plot illustrates the differentially expressed genes for each comparison between WT and ISM1^−/− lungs. A cutoff of 0.05 adjusted P value and twofold changes were used. Some significant DE genes were annotated. (H) Enriched GO biological processes of up-regulated genes. (I) Enriched GO biological processes of down-regulated genes. All the terms shown are enriched in ISM1^−/− lungs. (J)Adipoq expression comparing normal and asthmatic human lungs. RNA-Seq data were obtained and analyzed from the public databases Genotype-Tissue Expression B37 and B38_GC33 projects using Ingenuity Pathway Analysis software (OmicsSoft, Qiagen)