Journal: Frontiers in Pharmacology

Article Title: Guishaozichuan granules can attenuate asthma in rats via the MUC5AC/EGFR signaling pathway

doi: 10.3389/fphar.2022.1011751

Figure Lengend Snippet: Effects of GSZC granules on lung MUC5AC (A) , EGFR (B) mRNA expression in rats suffering from asthma. Values are the mean ± SD, n = 6 for each group. ▲▲ p < .01 versus the normal group, ** p < .01 versus the model group.

Article Snippet: MUC5AC antibody was purchased from Boster Group (20180824; Wuhan, China).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Guishaozichuan granules can attenuate asthma in rats via the MUC5AC/EGFR signaling pathway

doi: 10.3389/fphar.2022.1011751

Figure Lengend Snippet: Effects of GSZC granules on lung MUC5AC (A) , EGFR (B) protein expression in rats suffering from asthma. Values are the mean ± SD, n = 6 for each group. ▲▲ p < .01 versus the normal group, * p < .05, ** p < .01 versus the model group.

Article Snippet: MUC5AC antibody was purchased from Boster Group (20180824; Wuhan, China).

Techniques: Expressing

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: Pharmacological research

Article Title: S-allylmercaptocysteine inhibits mucin overexpression and inflammation via MAPKs and PI3K-Akt signaling pathways in acute respiratory distress syndrome.

doi: 10.1016/j.phrs.2020.105032

Figure Lengend Snippet: Fig. 2. Effects of SAMC on mucus hypersecre- tion induced by LPS in 16HBE cells and mice. (A) MUC5AC protein expression induced by LPS in 16HBE cells stimulated by different concentrations of LPS. (B) SAMC decreased the expression of MUC5AC protein both in vitro and in vivo. (C) SAMC reduced MUC5AC protein in mouse BALF determined by ELISA. (D) SAMC inhibited MUC5AC/ Muc5ac mRNA both in vitro and in vivo. (E) SAMC inhibited MUC5B/ Muc5b mRNA both in vitro and in vivo. Study groups: C: control group, orally treated with the vehicle only; M: model group, LPS was in- stilled into the lungs of mice through a high- pressure syringe; L and H: orally treated with SAMC 20 and 40 mg/kg after the LPS chal- lenge, respectively; P: positive control, NAC dosed at 500 mg/kg; n = 5 per group. * p < 0.05, ** p < 0.01, *** p < 0.001 compared with the control group; # p < 0.05, ## p < 0.01, ### p < 0.001 compared with the model group. The data are represented as means ± SD. All the experiments were re- peated three times.

Article Snippet: Mouse mucin-5 subtype AC (MUC5AC) ELISA kit was purchased from CUSABIO (Wuhan, China).

Techniques: Expressing, In Vitro, In Vivo, Enzyme-linked Immunosorbent Assay, Control, Positive Control

Journal: Pharmacological research

Article Title: S-allylmercaptocysteine inhibits mucin overexpression and inflammation via MAPKs and PI3K-Akt signaling pathways in acute respiratory distress syndrome.

doi: 10.1016/j.phrs.2020.105032

Figure Lengend Snippet: Fig. 3. Effects of SAMC on murine airway mucins. Sections show staining with AB-PAS (400×) and immunohistochemical staining (400×) of MUC5AC. The airway surface mucins were shown as the blue staining for the AB-PAS positive and brown staining for MUC5AC. Study groups: C: control group, orally treated with the vehicle only; M: model group, LPS was instilled into the lungs of mice through a high-pressure syringe; L and H: orally treated with SAMC at 20 and 40 mg/kg after the LPS challenge, respectively; P: positive control, NAC dosed at 500 mg/kg. n = 5 per group.

Article Snippet: Mouse mucin-5 subtype AC (MUC5AC) ELISA kit was purchased from CUSABIO (Wuhan, China).

Techniques: Staining, Immunohistochemical staining, Control, Positive Control

Journal: STAR Protocols

Article Title: Protocol for generating and characterizing a nasal epithelial model using imaging with application for respiratory viruses

doi: 10.1016/j.xpro.2024.103520

Figure Lengend Snippet: MUC5AC production by nasal epithelial culture after 7 days post air-liquid interface (ALI) Mucus was collected every 7 days during differentiation (mean of 3 wells per participant, ± SEM, are presented). 200 μL of PBS was added to the apical surface and incubated at 20°C–24°C for 5 min. MUC5AC ELISAs were performed to quantify mucus, indicating the presence of goblet cells (data from n = 3 tissue donors).

Article Snippet: Human MUC5AC (Mucin 5 Subtype AC) ELISA Kit , Biomatik , Cat# EKE61252.

Techniques: Incubation

Journal: STAR Protocols

Article Title: Protocol for generating and characterizing a nasal epithelial model using imaging with application for respiratory viruses

doi: 10.1016/j.xpro.2024.103520

Figure Lengend Snippet:

Article Snippet: Human MUC5AC (Mucin 5 Subtype AC) ELISA Kit , Biomatik , Cat# EKE61252.

Techniques: Virus, Control, Recombinant, Staining, Electron Microscopy, Sterility, Enzyme-linked Immunosorbent Assay, Gene Expression, Software, Microscopy, Membrane, Cell Culture, Pore Size, Marker, Adhesive, Low Protein Binding, Fluorescence, Inverted Microscopy

Journal: PLoS ONE

Article Title: β 2 -Adrenoceptor Involved in Smoking-Induced Airway Mucus Hypersecretion through β-Arrestin-Dependent Signaling

doi: 10.1371/journal.pone.0097788

Figure Lengend Snippet: Primer sequences used in experiments.

Article Snippet: Total mucin and MUC5AC level in BALF was assayed by use of rat mucin and MUC5AC ELISA kits (4A Biotech Co., Beijing, China).

Techniques:

Journal: PLoS ONE

Article Title: β 2 -Adrenoceptor Involved in Smoking-Induced Airway Mucus Hypersecretion through β-Arrestin-Dependent Signaling

doi: 10.1371/journal.pone.0097788

Figure Lengend Snippet: Cells were pre-incubated with 10 −5 M propranolol or 10 −6 M ICI118551 for 30 min before adding CSE (Pro+CSE, ICI+CSE) or were incubated with media (CON), CSE, 10 −5 M propranolol (Pro), or 10 −6 M ICI118551 (ICI) alone for 24 h. Quantitative RT-PCR and ELISA of MUC5AC mRNA (A) and protein level (B), respectively, and MUC5AC mRNA (C) and protein secretion (D) of cells transfected with Lipofectamine2000 (vehicle), a nonspecific control siRNA (NC-siRNA, 100 nM) or β 2 -AR–targeted siRNA (β 2 -siRNA, 100 nM) for 48 h before incubation with or without CSE for 24 h. Data are means ± SEM (n = 3).

Article Snippet: Total mucin and MUC5AC level in BALF was assayed by use of rat mucin and MUC5AC ELISA kits (4A Biotech Co., Beijing, China).

Techniques: Incubation, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Transfection, Control

Journal: PLoS ONE

Article Title: β 2 -Adrenoceptor Involved in Smoking-Induced Airway Mucus Hypersecretion through β-Arrestin-Dependent Signaling

doi: 10.1371/journal.pone.0097788

Figure Lengend Snippet: Western blot analysis of phospho-PKA (p-PKA) and total PKA (t-PKA) levels in cells preincubated with (A) Rp-8-Br-cAMPs or (B) H89 for 30 min before stimulation with or without CSE for 15 min. (C, D) RT-PCR analysis of MUC5AC mRNA expression after incubation with or without CSE for 24 h. Data are means ± SEM from 3 separate experiments. *** P <0.001 compared to CSE alone. NS, not significant.

Article Snippet: Total mucin and MUC5AC level in BALF was assayed by use of rat mucin and MUC5AC ELISA kits (4A Biotech Co., Beijing, China).

Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction, Expressing, Incubation

Journal: PLoS ONE

Article Title: β 2 -Adrenoceptor Involved in Smoking-Induced Airway Mucus Hypersecretion through β-Arrestin-Dependent Signaling

doi: 10.1371/journal.pone.0097788

Figure Lengend Snippet: Cells were transfected with Lipofectamine2000 alone (vehicle), a nonspecific control siRNA (NC-siRNA, 100 nM), β-arrestin1–targeted siRNA (A1-siRNA, 100 nM) or β-arrestin2–targeted siRNA (A2-siRNA, 100 nM) for 48 h before incubation with or without CSE for 24 h. (A, B) Western blot analysis of β-arrestin1 and β-arrestin2 protein level to access the knockdown efficiency. (C, D) Quantitative RT-PCR analysis of MUC5AC mRNA expression. Data are means ± SEM (n = 3). NS, not significant.

Article Snippet: Total mucin and MUC5AC level in BALF was assayed by use of rat mucin and MUC5AC ELISA kits (4A Biotech Co., Beijing, China).

Techniques: Transfection, Control, Incubation, Western Blot, Knockdown, Quantitative RT-PCR, Expressing

Journal: PLoS ONE

Article Title: β 2 -Adrenoceptor Involved in Smoking-Induced Airway Mucus Hypersecretion through β-Arrestin-Dependent Signaling

doi: 10.1371/journal.pone.0097788

Figure Lengend Snippet: Western blot analysis of phospho-ERK1/2 (p-ERK1/2), total ERK1/2 (t-ERK1/2), phospho-p38MAPK (p- p38MAPK) and total p38MAPK (t-p38MAPK) in cells treated with Lipofectamine2000 (vehicle), nonspecific control siRNA (NC-siRNA, 100 nM) or β-arrestin2–targeted siRNA (A2-siRNA, 100 nM) before CSE stimulation as in . (A, B) Representative western blot is from 3 independent experiments. (C, D) Data are the means ± SEM of 3 separate experiments. Cells were pretreated with ERK1/2 inhibitor PD98059 (50 µM) or p38MAPK inhibitor (5 µM) for 60 min before CSE treatment. (E) Quantitative RT-PCR analysis of MUC5AC mRNA expression. Data are means ± SEM (n = 3). NS, not significant.

Article Snippet: Total mucin and MUC5AC level in BALF was assayed by use of rat mucin and MUC5AC ELISA kits (4A Biotech Co., Beijing, China).

Techniques: Western Blot, Control, Quantitative RT-PCR, Expressing

Journal: PLoS ONE

Article Title: β 2 -Adrenoceptor Involved in Smoking-Induced Airway Mucus Hypersecretion through β-Arrestin-Dependent Signaling

doi: 10.1371/journal.pone.0097788

Figure Lengend Snippet: (A) MUC5AC glycoprotein expression in lung tissue by immunohistochemical staining. Bar = 200 µm and 100 µm in the upper and lower images, respectively. (B) Quantitative RT-PCR analysis of mRNA level of MUC5AC in lung homogenate. (C) ELISA of MUC5AC protein in BALF. Data are means ± SEM in each group.

Article Snippet: Total mucin and MUC5AC level in BALF was assayed by use of rat mucin and MUC5AC ELISA kits (4A Biotech Co., Beijing, China).

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

Journal: PLoS ONE

Article Title: β 2 -Adrenoceptor Involved in Smoking-Induced Airway Mucus Hypersecretion through β-Arrestin-Dependent Signaling

doi: 10.1371/journal.pone.0097788

Figure Lengend Snippet: Wtih CSE stimulation, β 2 -AR is activated, which increases cAMP and recruits many β -arrestins. Increased cAMP binds and activates PKA but does not stimulate MUC5AC transcription and production. β -arrestin2 phosphorylates ERK1/2 and p38MAPK, the key players, which activate transcription factors to stimulate MUC5AC transcription and production. β-AR antagonists, kinase inhibitors and molecular biological methods of interfering with signaling molecules are also shown.

Article Snippet: Total mucin and MUC5AC level in BALF was assayed by use of rat mucin and MUC5AC ELISA kits (4A Biotech Co., Beijing, China).

Techniques: