Journal: Cells

Article Title: MicroRNAs Associated with Chronic Mucus Hypersecretion in COPD Are Involved in Fibroblast-Epithelium Crosstalk.

doi: 10.3390/cells11030526

Figure Lengend Snippet: Figure 4. Effect of transfection with miR-708-5p mimic on FOXA2 expression and MUC5AC secretion by A549 epithelial cells. A549 cells were seeded on 24-well plates and transfected with miR-708-5p mimics or negative control siRNA (1 nM) in A549 cells in serum free medium. Cells were harvested for RNA isolation and cell-free supernatants were collected after 24 h. (a) Expression of FOXA2 was normalized to the housekeeping genes PPIA and B2M and compared between treatment with oligo control and miR-708-5p mimics. (b) MUC5AC levels were measured in cell-free supernatants by ELISA and compared between treatment with oligo control and miR-708-5p mimics. Four experiments were performed independently. Significant differences were determined by paired Student’s t-test. * p < 0.05 between the indicated values.

Article Snippet: Mucin 5AC (MUC5AC) levels were determined in supernatants using C96 Maxisorp NUNC Immuno-plate (Sigma-Aldrich, Darmstadt, Germany) coated with 100 μL/well of 500 ng/mL MUC5AC antibody (Thermo Fisher Scientific, Waltham, MA, USA) diluted in PBS (Gibco, California, CA, USA) overnight at room temperature on a platform shaker.

Techniques: Transfection, Expressing, Negative Control, Isolation, Control, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Serum MUC5AC protein levels are correlated with the development and severity of connective tissue disease-associated pulmonary interstitial lesions

doi: 10.3389/fimmu.2022.987723

Figure Lengend Snippet: Elevation of MUC5AC levels in CTD-ILDs (A). CTD-ILD, CTD-non ILD, and healthy controls. (B) . pSS-ILD, pSS-non ILD, and healthy controls. (C). SSc-ILD, SSc-non ILD, and healthy controls. (D) . DM/PM-ILD, DM/PM-non ILD, and healthy controls. Dot plots depict levels (measured by standard solid-phase enzyme-linked immunosorbent assay) of MUC5AC in individual serum samples from patients without ILD (interstitial lung abnormality ILD score 0), patients with ILD (indeterminate ILD (ILD score 1), mild ILD (ILD score 2), and advanced ILD (ILD score 3)) and healthy controls, Each symbol represents an individual patient; horizontal lines show the mean. p values were determined by Mann-Whitney U test.

Article Snippet: Serum MUC5AC and MUC5B protein levels were measured with double-antibody sandwich ELISA (Novus Biologicals, USA; NBP2-76703 and NBP2-76705) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Serum MUC5AC protein levels are correlated with the development and severity of connective tissue disease-associated pulmonary interstitial lesions

doi: 10.3389/fimmu.2022.987723

Figure Lengend Snippet: Elevation of MUC5B levels in CTD-ILDs (A) . CTD-ILD, CTD-non ILD, and healthy controls. (B). pSS-ILD, pSS-non ILD, and healthy controls. (C). SSc-ILD, SSc-non ILD, and healthy controls. (D). DM/PM-ILD, DM/PM-non ILD, and healthy control. The methodology used for these experiments was identical to those described in the legend for Figure 1 .

Article Snippet: Serum MUC5AC and MUC5B protein levels were measured with double-antibody sandwich ELISA (Novus Biologicals, USA; NBP2-76703 and NBP2-76705) according to the manufacturer’s instructions.

Techniques: Control

Journal: Frontiers in Immunology

Article Title: Serum MUC5AC protein levels are correlated with the development and severity of connective tissue disease-associated pulmonary interstitial lesions

doi: 10.3389/fimmu.2022.987723

Figure Lengend Snippet: Correlation between serum MUC5AC levels and ILD severity in patients with various autoimmune disease(s) (A) CTD-ILD. (B). pSS-ILD. (C). SSc-ILD. (D). DM/PM-ILD. Dot plot depicts the correlation between the level of MUC5AC and the severity of ILD in individual serum samples from patients with no ILD (interstitial lung abnormality (ILD score 0), uncertain ILD (ILD score 1), mild ILD (ILD score 2), or advanced ILD (ILD score 3) (measured by standard solid-phase enzyme-linked immunosorbent assay). Each symbol represents an individual patient; horizontal lines show the mean. Categorical variables were tested by using the Chi-square test and Fisher’s exact test. Statistical significance was determined using two-tailed tests and p<0.05.

Article Snippet: Serum MUC5AC and MUC5B protein levels were measured with double-antibody sandwich ELISA (Novus Biologicals, USA; NBP2-76703 and NBP2-76705) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Serum MUC5AC protein levels are correlated with the development and severity of connective tissue disease-associated pulmonary interstitial lesions

doi: 10.3389/fimmu.2022.987723

Figure Lengend Snippet: Correlation between serum MUC5B levels and ILD severity in patients with various autoimmune disease(s) (A) . CTD-ILD. (B). pSS-ILD. (C). SSc-ILD. (D) . DM/PM-ILD. The methodology used for these experiments was identical to those described in the legend for Figure 3 .

Article Snippet: Serum MUC5AC and MUC5B protein levels were measured with double-antibody sandwich ELISA (Novus Biologicals, USA; NBP2-76703 and NBP2-76705) according to the manufacturer’s instructions.

Techniques:

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: GPR120/FFAR4 stimulation attenuates airway remodeling and suppresses IL-4- and IL-13-induced airway epithelial injury via inhibition of STAT6 and Akt.

doi: 10.1016/j.biopha.2023.115774

Figure Lengend Snippet: Fig. 1. GPR120 agonist alleviates allergic airway inflammation and mucin secretion in OVA-induced asthmatic mice. Schematic overview of OVA-induced asthma experiment. Mice were sensitized and challenged with OVA and were intraperitoneally treated with vehicle, 20 mg/kg/day of GSK137647A, a GPR120 agonist or 5 mg/kg/day of dexamethasone (DEX). (B) Hematoxylin and eosin (H&E) staining of lung tissue sections. Infiltration of inflammatory cells in peribronchiolar regions (arrowheads). Scale bar: 100 µm. (C) Histopathological scores of peribronchiolar inflammatory cell infiltration. (D) Inflammatory cell counts in the bronchoalveolar lavage fluid (BALF). (E) Spleen to body weight ratio. (F) Mucin production in bronchiolar epithelial goblet cells stained with Periodic Acid-Schiff (PAS). Scale bar: 100 µm. (G) Histopathological scores of PAS-stained mucin production in bronchiolar epithelial goblet cells. (H) Levels of MUC5AC secretion in BALF measured by ELISA. (I) Representative Western blot analysis images of GPR120 protein expression in lung tissue samples (3 independent sets of experiment). (J) Densitometry values of GPR120 expression in the lung tissue samples. (n = 3–6 per condition; *P < 0.05; **P < 0.01, compared with control. #P < 0.05; ##P < 0.01; ###P < 0.001, compared with OVA-challenged group; ANOVA with Bonferroni multiple comparison test.).

Article Snippet: MUC5AC levels in BALF were determined by ELISA (CSB-E11040m; Cusabio, Houston, TX, USA), according to the manufacturer’s instructions.

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Control, Comparison

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: GPR120/FFAR4 stimulation attenuates airway remodeling and suppresses IL-4- and IL-13-induced airway epithelial injury via inhibition of STAT6 and Akt.

doi: 10.1016/j.biopha.2023.115774

Figure Lengend Snippet: Fig. 2. GPR120 inhibits IL-4- and IL-13-induced production of MUC5AC and cytokines in 16HBE human bronchial epithelial cells. 16HBE monolayers were incu bated with 10 ng/mL each of IL-4 and IL-13 and co-treated with or without 1 µM of GSK137647A for 48 h. To confirm activation of GPR120, cells were pretreated with 100 µM of AH7614, GPR120 antagonist for 1 h before the experiment. (A) Representative Western blot analysis images of GPR120 expression (3 independent sets of experiment). (B) Expression of MUC5AC (green) as depicted by immunofluorescent staining. Scale bar: 50 µm. (C) Fluorescent intensity ratio (MUC5AC to nuclei). (D) mRNA expression of MUC5AC. (E) Expression of cytokine transcripts IL6, IL8, IL25, and TSLP. (ns, non-significant difference; *P < 0.05; **P < 0.01, ***P < 0.001; ANOVA with Bonferroni multiple comparison test. n = 6–7 per condition.).

Article Snippet: MUC5AC levels in BALF were determined by ELISA (CSB-E11040m; Cusabio, Houston, TX, USA), according to the manufacturer’s instructions.

Techniques: Activation Assay, Western Blot, Expressing, Staining, Comparison

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: GPR120/FFAR4 stimulation attenuates airway remodeling and suppresses IL-4- and IL-13-induced airway epithelial injury via inhibition of STAT6 and Akt.

doi: 10.1016/j.biopha.2023.115774

Figure Lengend Snippet: Fig. 7. Schematic diagram demonstrating mechanisms by which GPR120 stimulation suppresses cytokine-induced airway remodeling in asthmatic models. Acti vation of type-I IL-4R (IL4Rα/γ-chain dimerization) emanates signaling via activation of insulin receptor substrate (IRS)-phosphoinositide 3-kinase (PI3K)-Akt pathway and JAK-STAT6 pathway, while type-II IL-4R (IL-4Rα/IL-13Rα1 dimerization) activates JAK-STAT6 pathway. GPR120 stimulation by GSK137647A at tenuates airway epithelial remodeling hallmarks including increased MUC5AC, tight junction disruption, and upregulated fibrotic markers. GPR120 stimulation attenuates the epithelial disrupting effects of IL-4 and IL-13 signaling via inhibition of STAT6 and Akt.

Article Snippet: MUC5AC levels in BALF were determined by ELISA (CSB-E11040m; Cusabio, Houston, TX, USA), according to the manufacturer’s instructions.

Techniques: Activation Assay, Disruption, Inhibition