Fig. 5

CS exposure activates reactive oxygen species (ROS) signaling downstream of miR-23a-5p/RAGE. A Principal component analysis for the expression of genes in different populations of 16-HBE cells; B the number of the differentially expressed genes (DEGs) in 16-HBE cells after different treatment; C KEGG pathway enrichment analysis of DEGs in the CSE group compared to controls; D Venn diagram of DEGs in different groups; E boxplots of the distribution of different groups of differential genes in HBE cells; F KEGG pathway enrichment analysis of downregulation-DEGs related with siRAGE and miR-23a-5p mimics treatment; G: WGCNA analysis was used to analyze the correlation between different treatments and DEGs in 16-HBE cells. H KEGG pathway enrichment analysis of strongly correlated genes in WGCNA analysis; I Immunofluorescence was used to detect the production of reactive oxygen species (ROS) after transfection of siRAGE and miR-23a-5p mimics in 16-HBE cells. J ROS positive staining was detected by immunofluorescence in alveolar lavage fluid cells from different groups of mice; K, L WB was used to detect levels of AKR protein in lung tissue of mice. M Immunofluorescence was used to detect ROS production in 16-HBE cells transfected with RAGE over-expression plasmids and ROS scavengers. N AGER mRNA levels were detected by rt-PCR after 16-HBE cells were transfected with a RAGE over-expression (OE) plasmids. O–Q mRNA levels of IL-6, IL-1β and TNF-α were detected by rt-PCR after 16-HBE cells transfected with RAGE OE plasmids. The experimental results were independently repeated three or more times, and Data are least squares means ± standard errors. *P < 0.05, **P < 0.01, ***p < 0.001