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: Molecular Medicine Reports

Article Title: Molecular mechanism underlying miR-130b-Sp1 transcriptional regulation in LPS-induced upregulation of MUC5AC in the bile duct epithelium

doi: 10.3892/mmr.2020.11745

Figure Lengend Snippet: Detection of MUC5AC expression in normal bile duct tissue and in tissues from patients with hepatoliths. (A) Immunohistochemistry assay of MUC5AC staining in paraffin-embedded sections from normal human subjects and patients with hepatoliths at different magnifications and quantitative analysis of MUC5AC expression level of each section. (B) Co-staining immunofluorescence experiments for MUC5AC and Sp1 expression in sections from normal human subjects and patients with hepatoliths (magnification, ×400), with quantitative analysis of Sp1 expression levels in each section. (C) Reverse transcription-quantitative PCR experiments for miR-130b expression in clinical tissue from normal human subjects and patients with hepatoliths. n=8 in the control group; n=10 in the hepatolith group. **P<0.01. MUC5AC, mucin 5AC; Sp1, specificity protein 1; miR, micro130b RNA.

Article Snippet: Subsequently, 250 μl concentrated cell supernatant solution was obtained from the control, LPS exposure, Sp1 shRNA transfection and miR130b mimics intervention groups and assayed using the human MUC5AC ELISA assay kit (cat. no. CSB-E10109h; Cusabio Technology, LLC) to determine MU5AC concentration.

Techniques: Expressing, Immunohistochemistry, Staining, Immunofluorescence, Reverse Transcription, Real-time Polymerase Chain Reaction, Control

Journal: Molecular Medicine Reports

Article Title: Molecular mechanism underlying miR-130b-Sp1 transcriptional regulation in LPS-induced upregulation of MUC5AC in the bile duct epithelium

doi: 10.3892/mmr.2020.11745

Figure Lengend Snippet: Detection of miR-130b, Sp1 and MUC5AC expression in HIBEpiCs following LPS treatment. (A) ELISA was performed to detect changes in MUC5AC expression in HIBEpiC supernatant. (B) RT-qPCR was performed to detect changes in MUC5AC and Sp1 mRNA expression in HIBEpiCs. (C) Western blotting was performed to detect changes Sp1 protein expression in HIBEpiCs. (D) RT-qPCR was performed to detect changes in miR-130b expression in HIBEpiCs. (E) immunofluorescence experiments for HIBEpiCs were performed to detect the transmembrane location of Sp1 in the control group compared with the 10 µg/ml LPS treatment group. Magnification, ×40. *P<0.05, **P<0.01. MUC5AC, mucin 5AC; Sp1, specificity protein 1; miR, microRNA; HIBEpiCs, human intrahepatic biliary epithelial cells; LPS, lipopolysaccharide; RT-qPCR, reverse transcription-quantitative PCR.

Article Snippet: Subsequently, 250 μl concentrated cell supernatant solution was obtained from the control, LPS exposure, Sp1 shRNA transfection and miR130b mimics intervention groups and assayed using the human MUC5AC ELISA assay kit (cat. no. CSB-E10109h; Cusabio Technology, LLC) to determine MU5AC concentration.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot, Immunofluorescence, Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Molecular mechanism underlying miR-130b-Sp1 transcriptional regulation in LPS-induced upregulation of MUC5AC in the bile duct epithelium

doi: 10.3892/mmr.2020.11745

Figure Lengend Snippet: MUC5AC, Sp1 and miR-130b levels in an intrahepatic bile duct stone animal model. (A) Sprague-Dawley rats underwent laparotomy and a PE tube was inserted into their common bile duct and fixed. The PE tube was placed from the back of the neck through a subcutaneous tunnel and fixed. (B) Days of each drug injection. (C) Rat serum levels of AST, ALT and TB were measured. (D) Images of rat bile smears after modeling. (E) Western blotting was performed to measure Sp1 expression in bile duct tissues from different groups. (F) RT-qPCR assay to detect expression of miR-130b in bile duct tissues across different groups. (G) RT-qPCR assay to detect expression levels of Sp1 and MUC5AC in bile duct tissues from different groups. (H) Correlation analysis of Sp1 and miR-130b mRNA expression levels. (I) Correlation analysis of MUC5AC and Sp1 mRNA expression levels. *P<0.05, **P<0.01 and ***P<0.0001. AST, aspartate transaminase; ALT, alanine aminotransferase; TB, total bilirubin; MUC5AC, mucin 5AC; Sp1, specificity protein 1; miR, microRNA; RT-qPCR, reverse transcription-quantitative PCR; PE, polyethylene.

Article Snippet: Subsequently, 250 μl concentrated cell supernatant solution was obtained from the control, LPS exposure, Sp1 shRNA transfection and miR130b mimics intervention groups and assayed using the human MUC5AC ELISA assay kit (cat. no. CSB-E10109h; Cusabio Technology, LLC) to determine MU5AC concentration.

Techniques: Animal Model, Injection, Western Blot, Expressing, Quantitative RT-PCR, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Molecular mechanism underlying miR-130b-Sp1 transcriptional regulation in LPS-induced upregulation of MUC5AC in the bile duct epithelium

doi: 10.3892/mmr.2020.11745

Figure Lengend Snippet: Identifying the direct binding position of Sp1 to the MUC5AC promoter sequence and further verification of the regulatory effect of Sp1 by transfection. (A) An online database was used to predict the possible binding positions for Sp1 to the MUC5AC promoter sequence. (B) Chromatin immunoprecipitation-qPCR experiments showing the Sp1 binding site on the MUC5AC promoter sequence and the alterations of Sp1 binding intensity between control and experimental groups (10 µg/ml LPS pretreatment for 24 h). HIBEpiCs were transfected with shRNA to overexpress Sp1 or shRNA to inhibit Sp1 or pretreated with 10 µg/ml mithramycin A (an Sp1 inhibitor). (C) RT-qPCR detection of Sp1 mRNA expression in HIBEpiCs. (D) Western blotting assay of Sp1 protein expression in HIBEpiCs. (E) RT-qPCR detection of MUC5AC mRNA expression in HIBEpiCs. (F) ELISA detection of MUC5AC expression in HIBEpiC supernatant. *P<0.05, **P<0.01 and ***P<0.0001. MUC5AC, mucin 5AC; Sp1, specificity protein 1; HIBEpiCs, human intrahepatic biliary epithelial cells; LPS, lipopolysaccharide; RT-qPCR, reverse transcription-quantitative PCR; shRNA, short hairpin RNA; NC, negative control; MA, mithramycin A.

Article Snippet: Subsequently, 250 μl concentrated cell supernatant solution was obtained from the control, LPS exposure, Sp1 shRNA transfection and miR130b mimics intervention groups and assayed using the human MUC5AC ELISA assay kit (cat. no. CSB-E10109h; Cusabio Technology, LLC) to determine MU5AC concentration.

Techniques: Binding Assay, Sequencing, Transfection, Chromatin Immunoprecipitation, Control, shRNA, Quantitative RT-PCR, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Negative Control

Journal: Molecular Medicine Reports

Article Title: Molecular mechanism underlying miR-130b-Sp1 transcriptional regulation in LPS-induced upregulation of MUC5AC in the bile duct epithelium

doi: 10.3892/mmr.2020.11745

Figure Lengend Snippet: Investigation of the direct binding position between miR-130b and the 3′-UTR sequence of Sp1 and further verification of the regulatory effect of miR-130b on MUC5AC and Sp1 expression. (A) Online tools were used to predict the binding position of miR-130b to the 3′-UTR sequence of Sp1, and the corresponding luciferase primer for a natural plasmid and a mutated plasmid were established. (B and C) Luciferase gene detection for miR-130b and the Sp1 3′-UTR sequence. HIBEpiCs were transfected with miR-130b overexpression mimics or miR-130b inhibitors for 24 h. (D) RT-qPCR detected changes in miR-130b expression in HIBEpiCs. (E) RT-qPCR detected changes in of Sp1 mRNA expression in HIBEpiCs. (F) Western blotting detected changes Sp1 protein expression in HIBEpiCs. (G) RT-qPCR detected changes in of MUC5AC mRNA expression in HIBEpiCs. (H) ELISA detected changes in MUC5AC levels in HIBEpiC supernatant. *P<0.05, **P<0.01 and ***P<0.0001. MUC5AC, mucin 5AC; Sp1, specificity protein 1; miR, microRNA; HIBEpiCs, human intrahepatic biliary epithelial cells; RT-qPCR, reverse transcription-quantitative PCR; 3′-UTR, 3′-untranslated region; LPS, lipopolysaccharide; NC, negative control; mut, mutant; shRNA, short hairpin RNA; inb, inhibitor.

Article Snippet: Subsequently, 250 μl concentrated cell supernatant solution was obtained from the control, LPS exposure, Sp1 shRNA transfection and miR130b mimics intervention groups and assayed using the human MUC5AC ELISA assay kit (cat. no. CSB-E10109h; Cusabio Technology, LLC) to determine MU5AC concentration.

Techniques: Binding Assay, Sequencing, Expressing, Luciferase, Plasmid Preparation, Transfection, Over Expression, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Negative Control, Mutagenesis, shRNA

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

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