per2  (Proteintech)

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

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: Per2 −/− mice aggravates OVA-induced AHR, airway inflammation, mucus production and fibrosis in experimental asthma. A Schematic illustrating the genetic approach used to knockout of Per2 mice. B Schematic overview of experimental design for the wildtype and Per2 −/− mouse model treated with OVA or PBS. C - E qPCR analysis and Western blot analysis the expression of PER2 in wildtype and Per2 −/− mouse lung tissues ( n = 3). F Non-invasive assessment of pulmonary function in mice after treatment with methacholine ( n = 5). G - K Total BALF cells, differential inflammatory cell counts of the lung sections were performed by Giemsa staining ( n = 5). L Representative images of H&E, PAS, and Masson staining of lung tissues from mice, original magnification 200 ×. Scale bars, 50 μm. M – O Inflammation scores based on H&E staining, PAS score based on PAS staining, and Masson’ Trichrome stain of lung tissues from mice ( n = 5). All data are expressed as mean ± SEM. Statistical significance was determined using unpaired Student’s t-test for (C, E); two-way repeated ANOVA followed by turkey’s comparison test for ( F ); and one-way ANOVA followed by Tukey’s post-hoc test for ( G - K , M - O ). AHR, airway hyperresponsiveness. * P < 0.05, ** P < 0.01, *** P < 0.001, ns, not significant

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: Knock-Out, Western Blot, Expressing, Staining, Comparison

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: PER2 deficiency aggravates airway remodeling via Wnt/β-catenin signaling pathway in vivo. A - E Representative Western blot images and statistical plots of protein quantitative analysis of N-cadherin, E-cadherin, MMP-9, and Vimentin in wildtype and Per2 −/− in lung tissues of mice ( n = 3). F - H qPCR analysis of the expression of Tgfb1 , Mmp2 and Muc5ac in lung tissues of mice ( n = 3). I - L Representative immunohistochemical images and quantitative estimation of E-cadherin, N-cadherin, and α-SMA in lung tissues of mice, original magnification 200 ×. Scale bars, 50 μm. All data are expressed as mean ± SEM. Statistical significance was determined using one-way ANOVA followed by Tukey’s post-hoc test for ( B - H , J - L ). * P < 0.05, ** P < 0.01, *** P < 0.001, ns, not significant

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: In Vivo, Western Blot, Expressing, Immunohistochemical staining

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: PER2 deficiency aggravates airway remodeling via Wnt signaling pathway in vivo. A - B A KEGG enrichment analysis in the lung tissues of WT-OVA compared to Per2 −/− -OVA. C - D Gene expression of TGF-β signaling pathway and EMT was assessed by Gene set enrichment analysis (GSEA). E - I Representative Western blotting images and statistical plots of protein quantitative analysis of β-catenin, GSK-3β, and p-GSK-3β in protein extracted from the lung tissues of mice ( n = 3). All data are expressed as mean ± SEM. Statistical significance was determined using one-way ANOVA followed by Tukey’s post-hoc test for (F-I). * P < 0.05, ** P < 0.01, *** P < 0.001, ns, not significant

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: In Vivo, Gene Expression, Western Blot

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: PER2 overexpression attenuates TGF-β1-induced EMT and migration in bronchial epithelial cells. A - B Representative Western blotting images and quantitative estimation of PER2 expression in TGF-β1-induced BEAS-2B cells for different doses was analyzed. C qPCR analysis of the PER2 expression in TGF-β1(10 ng/mL)-induced BEAS-2B cells. D - E Representative IF staining of PER2 in BEAS-2B cells stimulated with TGF-β1, original magnification 200 ×. Scale bars, 100 μm. F - H The expression of PER2 was studied by using qPCR and Western blot analysis. I - L Representative Western blotting images and quantitative estimation of N-cadherin, E-cadherin, α-SMA expression in TGF-β1-induced BEAS-2B cells. M - O qPCR analysis of the expression of MMP2 , SNAI1 and SNAI2 in BEAS-2B cells. P - Q Representative images and quantitative analysis of cell migration rates showing changes in BEAS-2B cell migration as assessed by scratch assays, original magnification 100 ×. Scale bars, 100 μm. R - S Representative images and quantitative analysis of cell migration rates showing changes in BEAS-2B cell migration as assessed by transwell assays, original magnification 100 ×. Scale bars, 100 μm. Statistical significance was determined using one-way ANOVA followed by Tukey’s post-hoc test for ( B , F , H , J - O , Q , S ), and unpaired Student’s t-test for ( C , E ). All data are expressed as mean ± SEM ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, ns, not significant

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: Over Expression, Migration, Western Blot, Expressing, Staining

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: PER2 suppresses TGF-β1-induced EMT by inhibiting Wnt/β-catenin signaling activation. A A volcano plot showed Wnt signaling pathway and other circadian related genes (red, upregulated genes; blue, downregulated genes). B A The heatmap of differentially expressed genes. Results were based on 3 samples of RNA sequencing. C A KEGG enrichment analysis in the cultured BEAS-2B cells of PER2 OE + TGF-β1 compared to Vector + TGF-β1. D - H Representative Western blotting images and Statistical plots of protein quantitative analysis of β-catenin, GSK-3β, and p-GSK-3β in TGF-β1-induced BEAS-2B cells. I - L Representative Western blotting images and quantitative analysis of the expression levels of the N-cadherin, β-catenin, α-SMA following intervention with the β-catenin agonist SKL2001. M - O The effect of SKL2001 on the protein content of β-catenin was detected by Western blot in the cytosol and nuclear. P - Q The protein expression of β-catenin in nuclear and its quantification were identified by Immunofluorescence, original magnification 200 ×. Scale bars, 100 μm. All data are expressed as mean ± SEM ( n = 3). Statistical significance was determined using one-way ANOVA followed by Tukey’s post-hoc test for ( E - H , J - L , N , O , Q ). * P < 0.05, ** P < 0.01, *** P < 0.001, ns, not significant

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: Activation Assay, RNA Sequencing, Cell Culture, Plasmid Preparation, Western Blot, Expressing, Immunofluorescence

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: Immunofluorescence results of PER2 suppresses TGF-β1-induced EMT by inhibiting Wnt/β-catenin signaling activation. A - C The immunofluorescence analysis of N-cadherin, E-cadherin, and α-SMA expression after expression of PER2, original magnification 200 ×. Scale bars, 100 μm. D - F The relative fluorescence intensity of N-cadherin, E-cadherin, and α-SMA. All data are expressed as mean ± SEM ( n = 3). Statistical significance was determined using one-way ANOVA followed by Tukey’s post-hoc test for ( D - F ). * P < 0.05, ** P < 0.01, *** P < 0.001, ns, not significant

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: Immunofluorescence, Activation Assay, Expressing, Fluorescence

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: Melatonin exerts its protective effects against EMT of asthma by upregulating the PER2. A , Chemical Structure of melatonin is shown. B , Establishment of mouse models of OVA-induced asthma and melatonin treatment. C , The mRNA expression level of Per2 after treatment with melatonin in the lung tissues of mice was analyzed by using qPCR ( n = 3). D-E , Representative Western blotting images and quantitative estimation of PER2 after treatment with melatonin in wildtype and Per2 -/- mouse lung tissues ( n = 3). F , Non-invasive assessment of pulmonary function in mice after treatment with melatonin ( n = 5). G-K , Total BALF cells and differential inflammatory cell counts were performed by Giemsa staining ( n = 5). L-N , Representative lung sections and semiquantitative analysis of peri-airway inflammatory infiltration, and goblet cell hyperplasia in OVA-exposed mice after melatonin administration, original magnification×100. Scale bars, 50 μm. O-R , Representative Western blotting images and quantitative estimation of N-cadherin, E-cadherin, and Vimentin in the lung tissues of mice ( n = 3). All data are expressed as mean ± SEM. Statistical significance was determined using one-way ANOVA followed by Tukey’s post-hoc test for ( C , E , G - K , M , N , P - R ); and two-way repeated ANOVA followed by turkey’s comparison test for (F). * P < 0.05, ** P < 0.01, *** P < 0.001, ns, not significant

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: Expressing, Western Blot, Staining, Comparison

Journal: Scientific Reports

Article Title: Circadian rhythms in pediatric high-grade gliomas may contribute to treatment efficacy

doi: 10.1038/s41598-025-17461-9

Figure Lengend Snippet: Circadian gene expression varies between pHGGs and pLGGs. ( A ) Interleaved scatter plots represent bulk RNA sequencing (rsem FPKM) analysis comparing circadian gene expression across pediatric low-grade glioma (pLGG, n = 426, blue) and pediatric high-grade glioma (pHGG, n = 211, red) samples from the Pediatric Brain Tumor Atlas (PBTA, Provisional). Data are expressed as mean ± SEM, and expression values are log₂-transformed FPKM counts. Statistical analysis was performed using a two-way ANOVA with Benjamini and Hochberg false discovery rate (FDR) to determine differences in gene expression between pHGGs and pLGGs. Each dot represents an individual tumor sample. Asterisks indicate statistically significant differences (* P < 0.05), while “NS” denotes non-significant comparisons. ( B ) Western blots of four representative pediatric high-grade glioma tissue samples (left) and four pediatric low-grade glioma tissue samples (right) comparing BMAL1, CLOCK, PER2, and REV-ERBα protein expression (sections of separate blots displayed; full blots are shown in Supplementary Figure S3). Of note, tissue sample HGG 3 is derived from the same patient as the STN-49 cell line. ( C ) Normalization to β-ACTIN control reveals BMAL1 ( p < 0.05, 1.57 vs. 0.98) and CLOCK ( p < 0.05, 0.98 vs. 0.39) protein expression is higher in high-grade gliomas compared to low-grade gliomas, REV-ERBα is lower ( p < 0.05, 0.12 vs. 0.55), and there are no significant differences in PER2 protein expression. Data are expressed as mean ± SEM.

Article Snippet: The following antibodies were used for immunoblotting: BMAL1 Rabbit mAb (A4714; ABclonal Technology, Woburn, MA), NR1D1 mAb (MA5-20772; Invitrogen), CLOCK anti-Rabbit (NB100-126; Novus Biologicals), PER2 anti-rabbit (NBP2-93587; Novus Biologicals), Beta-actin-HRP (MA5-15739-HRP; Invitrogen), anti-rabbit IgG HRP (7074; Cell Signaling Technology), and Goat anti-Mouse IgG Secondary Antibody, HRP (62-6520; Invitrogen).

Techniques: Gene Expression, RNA Sequencing, Expressing, Transformation Assay, Western Blot, Derivative Assay, Control