Journal: International Journal of Molecular Sciences

Article Title: Chr15q25 Genetic Variant rs16969968 Alters Cell Differentiation in Respiratory Epithelia

doi: 10.3390/ijms22136657

Figure Lengend Snippet: SNP rs16969968 is associated with a decrease in multiciliated cells in bronchi epithelia. ( A ) Representative micrographs showing the bronchial epithelia of α5WT- and α5SNP-expressing patients stained for haematoxylin and eosin. Magnification corresponding to the selected area is shown. ( B ) Box and whisker plots (median with min to max, the plus sign (+) represents the mean value) represent the measurements of epithelium height (µm), and the percentage of non-remodelled and remodelled (basal cell hyperplasia, goblet cell hyperplasia and metaplasia) epithelium in α5WT (black) and α5SNP-expressing (red) groups (n ≥ 7 for each group). ( C ) Representative micrographs showing the bronchial epithelia of α5WT and α5SNP-expressing patients stained for ciliated cells (Arl13b and FOXJ1, red), intermediate cells (uteroglobin, red), proliferative cells (Ki67, red), basal cells (P63, green), mucin-secreting cells (Muc5ac and Muc5b, green), intermediate filaments (vimentin, green) and cell nuclei (DAPI, blue). Magnification corresponding to the selected area is shown. ( D ) Box and whisker plots (median with min to max, the plus sign (+) represents the mean value) report the proportion of FOXJ1- and P63-positive cells per mm, the proliferative index, and the mean grey pixel values of mucin- and uteroglobin-associated fluorescence in α5WT (black) and α5SNP-expressing cells (red) (n ≥3 for each group). ** p < 0.01 α5WT vs. α5SNP.

Article Snippet: After PBS wash, a second primary antibody was used for 2 h at room temperature to highlight non-differentiated cells, secretory cells, and intermediate filaments on epithelia: p63 (AF1916, R&D Systems, Bio-Techne SAS, Noyal Châtillon sur Seiche, France), Muc5ac (NBP2-15196, Novus Biological, Bio-Techne SAS, Noyal Châtillon sur Seiche, France), Muc5b (E-AB-15988, ElabScience, CliniSciences SAS, Nanterre, France), and Vimentin (M0725, Agilent Dako, Les Ulis, France).

Techniques: Expressing, Staining, Whisker Assay, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Chr15q25 Genetic Variant rs16969968 Alters Cell Differentiation in Respiratory Epithelia

doi: 10.3390/ijms22136657

Figure Lengend Snippet: SNP rs16969968 alters respiratory airway epithelial cell differentiation. ( A ) Curves represent the TEER of AEC (n = 13) measured at ALI-1, ALI-7, ALI-14, ALI-21, ALI-28, and ALI-35 in α5WT (black) and α5SNP-expressing (red) cells. Means ± SEM are shown for each ALI time point. * p < 0.05; *** p < 0.001 α5WT vs. α5SNP. ( B ) Box and whisker plots (median with min to max, the plus sign (+) represents the mean value) represent the relative mRNA levels during ALI cultures by RT-qPCR ( n = 13) for differentiation markers (CK5, non-differentiated cells; FOXJ1, ciliated cells; MUC5AC/MUC5B, mucous-secreting cells) in α5SNP-expressing groups. * p < 0.05; ** p < 0.01; *** p < 0.001 α5WT vs. α5SNP. ( C ) Examples of micrographs taken from AEC cultures at ALI-14 and ALI-35 showing multiciliated (Arl13b, red) and mucin-secreting (Muc5ac, green) cells on the left, basal (CK13, red) and mucin-secreting (Muc5b, green) cells on the right. Nuclei are stained in blue (DAPI). Magnification corresponding to the selected area is shown. ( D ) Box and whisker plots (median with min to max, the plus sign (+) represents the mean value) represent the mean grey pixel values of Arl13b-, CK13-, muc5ac- and muc5b-associated fluorescence at ALI-14 and ALI-35 in α5WT (black) and α5SNP-expressing cells (red) (n ≥ 3 for each group). * p < 0.05 α5WT vs. α5SNP.

Article Snippet: After PBS wash, a second primary antibody was used for 2 h at room temperature to highlight non-differentiated cells, secretory cells, and intermediate filaments on epithelia: p63 (AF1916, R&D Systems, Bio-Techne SAS, Noyal Châtillon sur Seiche, France), Muc5ac (NBP2-15196, Novus Biological, Bio-Techne SAS, Noyal Châtillon sur Seiche, France), Muc5b (E-AB-15988, ElabScience, CliniSciences SAS, Nanterre, France), and Vimentin (M0725, Agilent Dako, Les Ulis, France).

Techniques: Cell Differentiation, Expressing, Whisker Assay, Quantitative RT-PCR, Staining, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Home Dust Mites Promote MUC5AC Hyper-Expression by Modulating the sNASP/TRAF6 Axis in the Airway Epithelium

doi: 10.3390/ijms23169405

Figure Lengend Snippet: HDM induces MUC5AC production in airway epithelial cells via TLR2/4-MyD88 signaling. ( A ) Expression of MUC5AC in 16HBE cells transfected with indicated siRNA and stimulated with HDM for 24 h. Results were normalized to the expression of ACTB (encoding β-actin) and control (siNC) cells. ( B ) Secretion of MUC5AC by 16HBE cells transduced as in ( A ) and stimulated with HDM. Data are representative of three independent experiments and presented as the mean ± SEM. ** p < 0.01.

Article Snippet: The collected BALF, lung tissue, or serum were assessed for expressions of IL-5, IL-13, and IgE levels using ELISA MAX Deluxe sets (Biolegend, San Diego, CA, USA) and a MUC5AC ELISA kit (Elabscience, Houston, TX, USA), according to the manufacturer’s instructions.

Techniques: Expressing, Transfection, Control

Journal: International Journal of Molecular Sciences

Article Title: Home Dust Mites Promote MUC5AC Hyper-Expression by Modulating the sNASP/TRAF6 Axis in the Airway Epithelium

doi: 10.3390/ijms23169405

Figure Lengend Snippet: Phosphorylation of sNASP regulates its interaction with TRAF6 and affects MUC5AC production following exposure to HDM. ( A ) The 16HBE cells were treated with HDM for different time periods and assessed by immunoblotting (IB) with antibodies against phosphorylated serine, NASP, or TRAF6 after immunoprecipitation (IP) with anti-NASP or by IB with anti-TRAF6 and anti-β-actin in total cell lysate (TCL). ( B ) RNA levels of MUC5AC in 16HBE cells transfected with the indicated siRNA and stimulated with HDM for 24 h. Results were normalized to the expression of ACTB (encoding β-actin) and control (siNC) cells. Data are presented as the mean ± SEM. ** p < 0.01 ( C ) Protein levels and ( D ) secretion of MUC5AC by 16HBE cells transduced as in ( B ) and stimulated with HDM. ( E ) The 16HBE cells were transfected with GFP-tagged wild-type (WT) sNASP or S158A and S164A mutants, followed by IB with antibody MUC5AC, GFP, or β-actin. Data are representative of three independent experiments.

Article Snippet: The collected BALF, lung tissue, or serum were assessed for expressions of IL-5, IL-13, and IgE levels using ELISA MAX Deluxe sets (Biolegend, San Diego, CA, USA) and a MUC5AC ELISA kit (Elabscience, Houston, TX, USA), according to the manufacturer’s instructions.

Techniques: Phospho-proteomics, Western Blot, Immunoprecipitation, Transfection, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Home Dust Mites Promote MUC5AC Hyper-Expression by Modulating the sNASP/TRAF6 Axis in the Airway Epithelium

doi: 10.3390/ijms23169405

Figure Lengend Snippet: The PEP-NASP peptide negatively regulates MUC5AC via NF-κB signaling. ( A ) The 16HBE cells were transfected with the siNC or siNASP, and then stimulated with HDM and assessed by an immunoblotting (IB) analysis with the indicated antibodies. ( B ) Luciferase activity in 16HBE cells transfected with a luciferase reporter vector driven by an NF-κB-responsive promoter, plus the siNC or NASP, and stimulated with HDM. Results were standardized to a control (siNC; set to 1). Data are the mean ± SEM for each group. ( C ) The 16HBE cells pretreated with the PEP-NASP or NASP were stimulated with HDM for and assessed by IB analysis with the indicated antibodies. ( D ) Luciferase activity of NF-κB ( E ) Secretion of MUC5AC by 16HBE cells transduced as described in ( C ). Data are representative of three independent experiments. * p < 0.05 and ** p < 0.01 (by a one-way ANOVA).

Article Snippet: The collected BALF, lung tissue, or serum were assessed for expressions of IL-5, IL-13, and IgE levels using ELISA MAX Deluxe sets (Biolegend, San Diego, CA, USA) and a MUC5AC ELISA kit (Elabscience, Houston, TX, USA), according to the manufacturer’s instructions.

Techniques: Transfection, Western Blot, Luciferase, Activity Assay, Plasmid Preparation, Control

Journal: International Journal of Molecular Sciences

Article Title: Home Dust Mites Promote MUC5AC Hyper-Expression by Modulating the sNASP/TRAF6 Axis in the Airway Epithelium

doi: 10.3390/ijms23169405

Figure Lengend Snippet: Effects of the PEP-NASP peptide on mRNA expression and secretion of inflammatory cytokines in BALF and lung tissues. ( A ) RNA expressions of IL-5, IL-13, and MUC5AC in lung tissues were determined by a RT-qPCR. ( B ) Levels of IL-5, IL-13, and MUC5AC in bronchoalveolar lavage fluid (BALF) were detected by an ELISA. n = 10, * p < 0.05 and ** p < 0.01 (by a one-way ANOVA).

Article Snippet: The collected BALF, lung tissue, or serum were assessed for expressions of IL-5, IL-13, and IgE levels using ELISA MAX Deluxe sets (Biolegend, San Diego, CA, USA) and a MUC5AC ELISA kit (Elabscience, Houston, TX, USA), according to the manufacturer’s instructions.

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Home Dust Mites Promote MUC5AC Hyper-Expression by Modulating the sNASP/TRAF6 Axis in the Airway Epithelium

doi: 10.3390/ijms23169405

Figure Lengend Snippet: Effects of the PEP-NASP peptide on the phosphorylation of sNASP in lung tissues. ( A ) Phospho-NASP-stained lung sections from each group were highlighted using immunohistochemical (IHC) staining by anti-pSerine158-sNASP. Scale bar = 100 μm ( B ) Lung from each group was analyzed by Western blotting with pSerine158-sNASP, sNASP, p-NF-κB p65, NF-κB p65, MUC5AC and β-actin antibodies. ( C ) Densitometric analysis of the Western blot shown below. Data represent the mean ± SEM of eight to twelve experiments (** p < 0.001 compared to PEP-treated control mice).

Article Snippet: The collected BALF, lung tissue, or serum were assessed for expressions of IL-5, IL-13, and IgE levels using ELISA MAX Deluxe sets (Biolegend, San Diego, CA, USA) and a MUC5AC ELISA kit (Elabscience, Houston, TX, USA), according to the manufacturer’s instructions.

Techniques: Phospho-proteomics, Staining, Immunohistochemical staining, Immunohistochemistry, Western Blot, Control

Journal: Frontiers in Oncology

Article Title: Prognostic significance of serum MUC5AC in resected pancreatic ductal adenocarcinoma: initial insights

doi: 10.3389/fonc.2025.1544928

Figure Lengend Snippet: Univariate logistic regression of serum MUC5AC on clinicopathological features.

Article Snippet: Serum MUC5AC samples were analyzed using the Human MUC5AC ELISA Kit (Catalog number NBP2-76703, Novus Biologicals, Centennial, CO) following the manufacturer’s instructions.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: Prognostic significance of serum MUC5AC in resected pancreatic ductal adenocarcinoma: initial insights

doi: 10.3389/fonc.2025.1544928

Figure Lengend Snippet: Serum MUC5AC (mean) distribution and clinicopathological features.

Article Snippet: Serum MUC5AC samples were analyzed using the Human MUC5AC ELISA Kit (Catalog number NBP2-76703, Novus Biologicals, Centennial, CO) following the manufacturer’s instructions.

Techniques:

Journal: Frontiers in Oncology

Article Title: Prognostic significance of serum MUC5AC in resected pancreatic ductal adenocarcinoma: initial insights

doi: 10.3389/fonc.2025.1544928

Figure Lengend Snippet: Univariate analysis for progression-free survival in neoadjuvant therapy group (n=21).

Article Snippet: Serum MUC5AC samples were analyzed using the Human MUC5AC ELISA Kit (Catalog number NBP2-76703, Novus Biologicals, Centennial, CO) following the manufacturer’s instructions.

Techniques: Biomarker Discovery

Journal: Frontiers in Oncology

Article Title: Prognostic significance of serum MUC5AC in resected pancreatic ductal adenocarcinoma: initial insights

doi: 10.3389/fonc.2025.1544928

Figure Lengend Snippet: Multivariate analysis for neoadjuvant therapy cohort.

Article Snippet: Serum MUC5AC samples were analyzed using the Human MUC5AC ELISA Kit (Catalog number NBP2-76703, Novus Biologicals, Centennial, CO) following the manufacturer’s instructions.

Techniques:

Journal: Frontiers in Oncology

Article Title: Prognostic significance of serum MUC5AC in resected pancreatic ductal adenocarcinoma: initial insights

doi: 10.3389/fonc.2025.1544928

Figure Lengend Snippet: Presurgery Multivariate analysis for survival.

Article Snippet: Serum MUC5AC samples were analyzed using the Human MUC5AC ELISA Kit (Catalog number NBP2-76703, Novus Biologicals, Centennial, CO) following the manufacturer’s instructions.

Techniques:

Journal: Frontiers in Oncology

Article Title: Prognostic significance of serum MUC5AC in resected pancreatic ductal adenocarcinoma: initial insights

doi: 10.3389/fonc.2025.1544928

Figure Lengend Snippet: Comparing low and high MUC5AC groups.

Article Snippet: Serum MUC5AC samples were analyzed using the Human MUC5AC ELISA Kit (Catalog number NBP2-76703, Novus Biologicals, Centennial, CO) following the manufacturer’s instructions.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: Prognostic significance of serum MUC5AC in resected pancreatic ductal adenocarcinoma: initial insights

doi: 10.3389/fonc.2025.1544928

Figure Lengend Snippet: The difference in survival between low and high serum MUC5AC groups in neoadjuvant therapy group (A, B) and FOLFIRINOX sub-group (C, D) .

Article Snippet: Serum MUC5AC samples were analyzed using the Human MUC5AC ELISA Kit (Catalog number NBP2-76703, Novus Biologicals, Centennial, CO) following the manufacturer’s instructions.

Techniques:

Journal: Frontiers in Oncology

Article Title: Prognostic significance of serum MUC5AC in resected pancreatic ductal adenocarcinoma: initial insights

doi: 10.3389/fonc.2025.1544928

Figure Lengend Snippet: Multivariate analysis of upfront surgery population for survival (n=17).

Article Snippet: Serum MUC5AC samples were analyzed using the Human MUC5AC ELISA Kit (Catalog number NBP2-76703, Novus Biologicals, Centennial, CO) following the manufacturer’s instructions.

Techniques:

Journal: Frontiers in Oncology

Article Title: Prognostic significance of serum MUC5AC in resected pancreatic ductal adenocarcinoma: initial insights

doi: 10.3389/fonc.2025.1544928

Figure Lengend Snippet: The difference in progression-free survival (A) and overall survival (B) between low and high serum MUC5AC groups in upfront surgery group.

Article Snippet: Serum MUC5AC samples were analyzed using the Human MUC5AC ELISA Kit (Catalog number NBP2-76703, Novus Biologicals, Centennial, CO) following the manufacturer’s instructions.

Techniques:

Journal: Frontiers in Oncology

Article Title: Prognostic significance of serum MUC5AC in resected pancreatic ductal adenocarcinoma: initial insights

doi: 10.3389/fonc.2025.1544928

Figure Lengend Snippet: Predicting recurrence using MUC5AC and CA19-9 post-surgery.

Article Snippet: Serum MUC5AC samples were analyzed using the Human MUC5AC ELISA Kit (Catalog number NBP2-76703, Novus Biologicals, Centennial, CO) following the manufacturer’s instructions.

Techniques: