Journal: Respiratory Research
Article Title: The circadian clock component PER2 deficiency aggravates airway epithelial remodeling through Wnt/β-catenin signaling pathway
doi: 10.1186/s12931-026-03522-8
Figure Lengend Snippet: Integrative bioinformatics and experimental validation identify circadian clock component PER2 as a key regulator in asthma. A Venn diagram of significant genes from GSE63142 and CRGs. B The Random Forest algorithm was selected 11 genes based on Mean Decrease Accuracy scores greater than 2.0. C The LASSO logistic regression algorithm determined 16 genes. D The PPI network analysis identified top 10 genes by the CytoHubba plugin. E Venn diagram of the overlapping hub genes between the Random Forest, LASSO algorithm and PPI network. F - G Association of PER2 and PER3 mRNA expression levels in bronchial epithelial cells between control ( n = 27), mild-moderate asthma (MMA) ( n = 72), and severe asthma (SA) ( n = 56) patients from GSE63142 . PER2 and PER3 mRNA expression levels have been logarithm base 2 (log2)-transformed. H - I Association of PER2 and PER3 mRNA expression levels in bronchial epithelial brushings between control ( n = 30), and patients with asthma ( n = 51) from GSE41861 . PER2 and PER3 mRNA expression levels have been log2-transformed. J - K Association of PER2 and PER3 mRNA expression levels in bronchial epithelial cells between control ( n = 20), mild-moderate asthma (MMA) ( n = 50), and severe asthma (SA) ( n = 38) patients from GSE43696 . PER2 and PER3 mRNA expression levels have been log2-transformed. L - O Correlation between PER2 mRNA expression in primary bronchial epithelial cells and FEV 1 (L), FEV 1 % predicted, FVC (L), IgE (IU/mL) was determined by Spearman analysis in patients with asthma from GSE201955 ( n = 79). P qPCR analysis of the expression of Per2 in lung tissues of mice ( n = 3). Q - R Western blot analysis the expression of PER2 in lung tissues of mice treated with OVA or PBS ( n = 3). S - T Representative Immunofluorescence staining of PER2 (Red) in the airway epithelium (SCGB1A1 Green) from asthmatic and control mice ( n = 3), original magnification 200 ×. Scale bars, 100 μm. Statistical significance was determined using the Kruskal-Wallis test followed by turkey’s comparison test for ( F , G , J , K ); Mann-Whitney U test for ( H , I ); spearman correlation for ( L - O ); and unpaired Student’s t-test for ( P , R , T ). CRG, circadian related genes; LASSO, least absolute shrinkage and selection operator; RF, random forest; PPI, protein-protein interaction; MMA, mild-moderate asthma; SA, severe asthma; FEV 1 , forced expiratory volume in 1 s; FVC, forced vital capacity; IgE, immunoglobulin E. ** P < 0.01, *** P < 0.001
Article Snippet: After blocking with 5% BSA at 37 °C for 30 min, sections were incubated overnight at 4 °C with anti-PER2 antibody (1:100, NB100-125, Novus) and anti-SCGB1A1/CC10 antibody (1:50, 26,909–1-AP, Proteintech).
Techniques: Biomarker Discovery, Expressing, Control, Transformation Assay, Western Blot, Immunofluorescence, Staining, Comparison, MANN-WHITNEY, Selection