Journal: Frontiers in Medicine

Article Title: TOLLIP and MUC5B single nucleotide polymorphisms among interstitial lung disease patients from Western India

doi: 10.3389/fmed.2026.1806788

Figure Lengend Snippet: The squared Pearson’s correlation coefficient R 2 (A) , and D prime (B) were used as a measure of LD between pairs of SNPs using Haploview. Low R 2 values were consistently observed for all the SNP pairs. The SNPs rs111521887 and rs5743894 had a R 2 value of 39 and a D’ value of 75, indicating no significant linkage. The 2 other pairs, with higher D’ values – rs35705950 and rs3750920, and the pair rs35705950 and rs5743890, show similar results with high D’ values but low R 2 . This pattern suggests that although recombination events between these loci may be limited (high D’), the allelic correlation is weak (low R 2 ).

Article Snippet: SNPs including MUC5B rs35705950, TOLLIP rs111521887, TOLLIP rs5743854, and TOLLIP rs5743890 were identified using Sanger’s sequencing method, amplified using locus-specific primers, and sequenced using the ABI 3730xl DNA analyzer (Applied Biosystems, USA).

Techniques:

Journal: Molecular Medicine Reports

Article Title: Cigarette smoke combined with Klebsiella pneumoniae induce damage to the air-blood barrier in chronic obstructive pulmonary disease rats via the MAPK/NF-κB/IκBα pathway

doi: 10.3892/mmr.2026.13797

Figure Lengend Snippet: Injury of mucus hypersecretion and oxidative damage in all groups. (A) AB-PAS staining images of the airway in all groups (AB-PAS; ×200 magnification and ×400 magnification). Arrows indicate goblet cells. (B) The expression level of MUC5AC and MUC5B in the airway tested using immunohistochemistry (×200 magnification). (C) The expression of IL-6, IL-10, TNF-α, T-SOD, MDA, MMP-9 and TIMP-1 in lung tissue and serum of rats in all groups. The values are expressed as mean ± SE; (n=8). *P<0.05 and **P<0.01 vs. normal group. AB-PAS, Alcian blue-periodic acid-schiff; MUC5AC, mucoprotein-5AC; MUC5B, mucoprotein-5B; T-SOD, total superoxide dismutase; MDA, malondialdehyde; MMP-9, matrix metalloproteinase 9; TIMP-1, tissue inhibitor of metalloproteinases 2; IOD, integrated optical density.

Article Snippet: Non-specific binding was blocked by incubating the slides in 5% BSA (cat. no. A8020; Beijing Solarbio Science & Technology Co., Ltd.) at 37°C for 1 h. After blocking, the sections were incubated overnight at 4°C with the following primary antibodies: MUC5AC (1:500; cat. no. 85868-1-RR; Proteintech Group, Inc.), MUC5B (1:500; cat. no. 28118-1-AP; Proteintech Group, Inc.), zonula occludens (ZO)-1 (1:200; cat. no. AF5145; Affinity Biosciences) and claudin-5 (1:200; cat. no. AF5216; Affinity Biosciences).

Techniques: Staining, Expressing, Immunohistochemistry

Journal: Frontiers in Immunology

Article Title: Identification of MUC5B as a lymph node metastasis-associated gene in lung adenocarcinoma through integrated transcriptomic and machine learning approaches

doi: 10.3389/fimmu.2025.1666240

Figure Lengend Snippet: Expression of MUC5B in tumor and normal tissues and its prognostic impact on overall survival in LUAD. (A) MUC5B expression in tumor and normal tissues; (B) Kaplan-Meier survival curve for MUC5B. ****P < 0.0001.

Article Snippet: After endogenous peroxidase blocking (3% H 2 O 2 ) and antigen retrieval (citrate buffer), sections were incubated with anti-MUC5B antibody (1:100, HUABIO) at 4°C overnight, followed by HRP-conjugated secondary antibody (30 min, RT).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: Identification of MUC5B as a lymph node metastasis-associated gene in lung adenocarcinoma through integrated transcriptomic and machine learning approaches

doi: 10.3389/fimmu.2025.1666240

Figure Lengend Snippet: Identification of key genes and functional analysis. (A) Intersection of co-expressed genes related to lymph node metastasis and MUC5B; (B) PPI network of 25 candidate genes; (C) Sub-network constructed with cytoHubba; (D) GO functional enrichment analysis of key genes; (E) KEGG pathway enrichment analysis of key genes.

Article Snippet: After endogenous peroxidase blocking (3% H 2 O 2 ) and antigen retrieval (citrate buffer), sections were incubated with anti-MUC5B antibody (1:100, HUABIO) at 4°C overnight, followed by HRP-conjugated secondary antibody (30 min, RT).

Techniques: Functional Assay, Construct

Journal: Frontiers in Immunology

Article Title: Identification of MUC5B as a lymph node metastasis-associated gene in lung adenocarcinoma through integrated transcriptomic and machine learning approaches

doi: 10.3389/fimmu.2025.1666240

Figure Lengend Snippet: Construction of gene features and correlation between key genes and immune cells. (A) LASSO coefficient profile of 23 core genes; (B) SVM-RFE result validation using 10-fold CV RMSE; (C) Venn diagram showing the intersection of LASSO and SVM-selected genes; (D) SHAP value scores of each input feature; (E) SHAP summary plot showing the impact of each feature on the full model output. (F) Heatmap showing the correlation between the model and immune scores; (G) miRNA-mRNA-TF network for MUC5B.

Article Snippet: After endogenous peroxidase blocking (3% H 2 O 2 ) and antigen retrieval (citrate buffer), sections were incubated with anti-MUC5B antibody (1:100, HUABIO) at 4°C overnight, followed by HRP-conjugated secondary antibody (30 min, RT).

Techniques: Biomarker Discovery

Journal: Frontiers in Immunology

Article Title: Identification of MUC5B as a lymph node metastasis-associated gene in lung adenocarcinoma through integrated transcriptomic and machine learning approaches

doi: 10.3389/fimmu.2025.1666240

Figure Lengend Snippet: Expression and survival analysis of MUC5B in human LUAD patients. (A, B) Immunohistochemistry for MUC5B in LUAD without LN metastasis (A) and with LN metastasis (B) . (C) Quantitative analysis of H-score of MUC5B expression in LN metastasis (-) and LN metastasis (+) patients (n=30 vs. 35). (D) Overall survival (OS) in the high and low MUC5B groups. (*P<0.05, H-Score = Σ (pi × i) = (% weak cells × 1) + (% moderate cells × 2) + (% strong cells × 3)).

Article Snippet: After endogenous peroxidase blocking (3% H 2 O 2 ) and antigen retrieval (citrate buffer), sections were incubated with anti-MUC5B antibody (1:100, HUABIO) at 4°C overnight, followed by HRP-conjugated secondary antibody (30 min, RT).

Techniques: Expressing, Immunohistochemistry

Journal: Frontiers in Immunology

Article Title: Identification of MUC5B as a lymph node metastasis-associated gene in lung adenocarcinoma through integrated transcriptomic and machine learning approaches

doi: 10.3389/fimmu.2025.1666240

Figure Lengend Snippet: MUC5B knockdown reduces proliferation, migration, invasion, and colony formation in LUAD cells. (A, B) CCK-8 assays demonstrating significantly reduced cell viability in A549 and H1975 cells transfected with siMUC5B compared to siNC controls over a 96-hour period. (C, D) qRT-PCR results confirming the effective knockdown of MUC5B mRNA expression in A549 and H1975 cells following siMUC5B transfection. (E) Representative images from the colony formation assay showing significantly fewer colonies in A549 and H1975 cells transfected with siMUC5B compared to siNC. (F) Quantification of colony formation results, revealing a marked reduction in colony numbers in MUC5B-knockdown cells. (G) Wound healing assay showing impaired migration in both A549 and H1975 cells 24 hours after MUC5B knockdown. (H) Quantification of the wound closure rate demonstrates a significant reduction in the migration capacity of siMUC5B-transfected cells compared to controls. (I) Transwell migration and invasion assays showing that MUC5B knockdown significantly reduces the migratory and invasive capabilities of A549 and H1975 cells. (J) Quantification of migration and invasion reveals a substantial decrease in cell movement and invasiveness in MUC5B-depleted cells. Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: After endogenous peroxidase blocking (3% H 2 O 2 ) and antigen retrieval (citrate buffer), sections were incubated with anti-MUC5B antibody (1:100, HUABIO) at 4°C overnight, followed by HRP-conjugated secondary antibody (30 min, RT).

Techniques: Knockdown, Migration, CCK-8 Assay, Transfection, Quantitative RT-PCR, Expressing, Colony Assay, Wound Healing Assay

Journal: Frontiers in Immunology

Article Title: Identification of MUC5B as a lymph node metastasis-associated gene in lung adenocarcinoma through integrated transcriptomic and machine learning approaches

doi: 10.3389/fimmu.2025.1666240

Figure Lengend Snippet: RNA-seq reveals transcriptomic changes following MUC5B knockdown in LUAD cells. (A) Volcano plot showing DEGs between si-MUC5B and si-NC groups. Red and blue dots represent significantly upregulated and downregulated genes (|log2FC| > 1, p < 0.05). (B) GO enrichment analysis of DEGs. (C) KEGG pathway analysis highlighting key pathways related to DNA replication and cell cycle. (D) Heatmap of the 13-gene model and GINS family expression after MUC5B knockdown. (E) qPCR validation showing reduced expression of GINS1, GINS2, GINS3 and GINS4 following MUC5B silencing in A549 cells. ***P < 0.001, ****P < 0.0001.

Article Snippet: After endogenous peroxidase blocking (3% H 2 O 2 ) and antigen retrieval (citrate buffer), sections were incubated with anti-MUC5B antibody (1:100, HUABIO) at 4°C overnight, followed by HRP-conjugated secondary antibody (30 min, RT).

Techniques: RNA Sequencing, Knockdown, Expressing, Biomarker Discovery