Research Paper Volume 15, Issue 7 pp 2395—2417

Figure 5. Epithelial and endothelial cell populations mediate p21-dependent inflammatory responses following chronic LPS inhalation. WT and p21^-/- mice were exposed to either PBS or aerosolized LPS (0.5 mg/ml), 3 times a week for 10 weeks. At 48 hours following the last LPS exposure, whole lungs were dissected and dissociated into single cell suspensions and analyzed by flow cytometry and sorting for subsequent RNA sequencing. Data was analyzed as follows: (A) Hierarchical clustering heatmap of differentially expressed genes (DEGs) in epithelial, endothelial and immune cells populations in the mice lungs. One sample of the immune cells population, in the PBS p21^-/- mice group, was excluded from the analysis. (B) Functional analysis of the DEGs in epithelial, endothelial and immune cells populations in the mice lungs. Abbreviations: antigen processing and presentation - antigen presentation, cytokine-mediated signaling pathway - cytokine-mediated signaling, external side of plasma membrane - external plasma membrane, glomerulus vasculature morphogenesis - glomerulus vasculature morphogenesis, positive regulation of immune response - immune response regulation, positive regulation of MAPK cascade - regulation of MAPK cascade, regulation of cytokine production - cytokine production, regulation of tumor necrosis factor superfamily cytokine production - TNF cytokine production. (C) Heatmap based on functional analysis of the upregulated DEGs in epithelial, endothelial and immune cells populations in the mice lungs. (D) Number of upregulated LPS responsive DEGs in epithelial, endothelial and immune cells populations in the mice lungs. (E) Number of upregulated LPS responsive DEGs in p21^-/- affected clusters in epithelial, endothelial and immune cells populations in the mice lungs. Data information: Data were analyzed using Chi-squared test. ***P<0.0005. (A–E), n=4-5 independent repeats.