mouse anti muc5ac monoclonal antibody  (Bio-Rad)

Journal: Journal of Experimental Pharmacology

Article Title: Discovery of Fungus-Derived Nornidulin as a Novel TMEM16A Inhibitor: A Potential Therapy to Inhibit Mucus Secretion in Asthma

doi: 10.2147/JEP.S427594

Figure Lengend Snippet: Effect of nornidulin on Ca 2+ -dependent mucin secretion in Calu-3 cells. ( A ) Immunofluorescence staining of MUC5AC. Hoechst staining was used to determine the cell area. MUC5AC, a marker of pathogenic mucin in asthma, was labeled to evaluate depletion of intracellular mucin store that indicates mucin secretion. Ionomycin treatment was performed to promote Ca 2+ -dependent mucin secretion. Nornidulin (10 µM) and a TMEM16A inhibitor MONNA were able to suppress ionomycin-induced depletion of intracellular mucin store, suggesting that nornidulin was capable of inhibiting mucin secretion. The scale bars of 50 µm were shown. ( B ) Analyses of intracellular mucin levels. The summary and statistical analyses of data are shown. Results were expressed as % of control ± S.E.M. (n = 4–6). ** p < 0.01 compared with control group (one-way ANOVA). # p < 0.05; ## p < 0.01 compared with ionomycin-treated group (one-way ANOVA).

Article Snippet: The cells were fixed with 4% paraformaldehyde and permeabilized with 0.05% Triton X before overnight incubation with mouse anti-MUC5AC monoclonal antibody (45M1) (catalog number: MA5-12178, Thermo Fisher Scientific Inc., Waltham, MA, USA).

Techniques: Immunofluorescence, Staining, Marker, Labeling

Journal: Journal of Experimental Pharmacology

Article Title: Discovery of Fungus-Derived Nornidulin as a Novel TMEM16A Inhibitor: A Potential Therapy to Inhibit Mucus Secretion in Asthma

doi: 10.2147/JEP.S427594

Figure Lengend Snippet: Effect of nornidulin in the treatment of inflammation-associated mucus hypersecretion in OVA-challenged mouse model of asthma. ( A ) Timeline and detailed protocol of OVA-challenged mouse model of asthma establishment. ( B ) Effect of nornidulin and dexamethasone (DEX) on percent spleen weight. ( C ) H&E and PAS staining of airway tissues from OVA-challenged mice treated with or without nornidulin. Dexamethasone (DEX) was used as a control of this experiment. The scale bars of 40 µm were shown with the red arrows and the black arrow heads indicating clusters of eosinophils and mucus-producing cells, respectively. ( D ) Inflammatory score and ( E ) % PAS-positive area of airway tissues from OVA-challenged mice treated with or without nornidulin. Dexamethasone (DEX) was used as a control of these experiment. ( F ) Effect of nornidulin on in vivo mucus hypersecretion in OVA-challenged mice. Mucus hypersecretion was evaluated by the levels of MUC5AC, a pathogenic mucin, collected from bronchoalveolar lavage fluid (BALF). Both nornidulin and dexamethasone (DEX) significantly reduced mucus hypersecretion. ( G ) Effect of nornidulin on total immune cell number. BALF was collected and immune cells were observed and counted under microscope. Dexamethasone (DEX), but not nornidulin, significantly reduced total cells in BALF. Results were analyzed from 5–7 independent experiments and shown as a means of control ± S.E.M. * p < 0.05; ** p < 0.01; *** p < 0.001 compared with control group (one-way ANOVA). # p < 0.05; ## p < 0.01; ### p < 0.001; NS, non-statistical difference compared with OVA-challenged groups without any treatment (one-way ANOVA).

Article Snippet: The cells were fixed with 4% paraformaldehyde and permeabilized with 0.05% Triton X before overnight incubation with mouse anti-MUC5AC monoclonal antibody (45M1) (catalog number: MA5-12178, Thermo Fisher Scientific Inc., Waltham, MA, USA).

Techniques: Staining, In Vivo, Microscopy