Journal: Scientific Reports

Article Title: Function-specific IL-17A and dexamethasone interactions in primary human airway epithelial cells

doi: 10.1038/s41598-022-15393-2

Figure Lengend Snippet: IL-17A induces gene expression that is insensitive to dexamethasone (Dex) in human airway epithelial cells (hAECs). hAECs were cultured and differentiated at an air liquid interface (ALI) and exposed to IL-17A (10 ng/ml) with or without dexamethasone (10 nM) for 14 days. Gene expression was determined by bulk RNA sequencing and differentially expressed genes were analyzed and presented as heatmap of the top 10 most regulated genes, with each column in the groups representing an individual donor ( a ) and by a volcano plot ( b ). Pathway analysis (GSEA) of the statistically significant genes was done and presented as the top 10 most regulated pathways ( c ). The IL-17A gene signature of the top 10 differentially expressed genes was calculated as Z-score ratio ( d ). Data represent means ± SEM. N = 4 donors.

Article Snippet: FDR q-values were assessed using the Gene Set Enrichment Analysis webtool, Broad Institute, UC San Diego ( https://www.gsea-msigdb.org/gsea/index.jsp ).

Techniques: Gene Expression, Cell Culture, RNA Sequencing

Journal: Scientific Reports

Article Title: Function-specific IL-17A and dexamethasone interactions in primary human airway epithelial cells

doi: 10.1038/s41598-022-15393-2

Figure Lengend Snippet: Dexamethasone (Dex)-inducible genes are dampened by the presence of IL-17A in human airway epithelial cells (hAECs). hAECs were cultured and differentiated at an air liquid interface (ALI) and exposed to IL-17A (10 ng/ml) with or dexamethasone (10 nM) for 14 days. Gene expression was determined by bulk RNA sequencing and differentially expressed genes were analyzed and presented as a heatmap of the top 10 most regulated genes by dexamethasone, with each column in the groups representing an individual donor ( a ) and by a volcano plot ( b ). Pathway analysis (GSEA) of the differentially expressed genes was done subsequently ( c ). The dexamethasone gene signature was calculated as Z-score ratio of the most regulated genes ( d ). Canonical corticosteroid gene expression, HSD11B2 ( e ) and FKBP5 ( f ), were analyzed from normalized gene counts. Data represent means ± SEM. N = 4 donors. Statistical analyses were performed with paired two-way ANOVA and Šidák’s multiple comparison test, *p < 0.05, ***p < 0.001.

Article Snippet: FDR q-values were assessed using the Gene Set Enrichment Analysis webtool, Broad Institute, UC San Diego ( https://www.gsea-msigdb.org/gsea/index.jsp ).

Techniques: Cell Culture, Gene Expression, RNA Sequencing, Comparison

Journal: Scientific Reports

Article Title: Function-specific IL-17A and dexamethasone interactions in primary human airway epithelial cells

doi: 10.1038/s41598-022-15393-2

Figure Lengend Snippet: Dexamethasone (Dex) reverses IL-17A-induced epithelial barrier disruption in human airway epithelial cells (hAECs). hAECs were cultured and differentiated at an air liquid interface (ALI) and exposed to IL-17A (10 ng/ml) with or without dexamethasone (10 nM) for 14 days. Epithelial barrier function was assessed by permeability to fluorescein isothiocyanate-dextran (FITC-dextran) after 2 weeks incubation with IL-17A or dexamethasone of combination of both. FITC concentration was measured and calculated from fluorescence intensity. Data are from N = 7 donors ( a ). Gene expression was determined by bulk RNA sequencing and differentially expressed genes were analyzed. Statistically significant genes were used for pathway analysis (GSEA) and presented as top 10 most regulated pathways ( b ). String analysis ( https://string-db.org/ ) was done to the genes that are related to cilia function and development ( c ). Data represent means ± SEM. N = 4 donors. Statistical analyses were performed with paired one-way ANOVA and Tukey’s multiple comparison test, *p < 0.05.

Article Snippet: FDR q-values were assessed using the Gene Set Enrichment Analysis webtool, Broad Institute, UC San Diego ( https://www.gsea-msigdb.org/gsea/index.jsp ).

Techniques: Disruption, Cell Culture, Permeability, Incubation, Concentration Assay, Fluorescence, Gene Expression, RNA Sequencing, Comparison