Journal: International Journal of Molecular Sciences
Article Title: Mucus Release and Airway Constriction by TMEM16A May Worsen Pathology in Inflammatory Lung Disease
Figure Lengend Snippet: Niclosamide inhibits expression of MUC5AC and SPDEF in Calu-3 cells. ( A ) Western blot indicating upregulation of TMEM16A in Calu-3 cells by IL-13 (20 ng/mL, 72 h) and inhibition of expression by niclosamide (1 µM). Blots were performed in triplicates. ( B ) RT-PCR analysis of the expression of MUC5AC, TMEM16A, and SAM pointed domain-containing ETS transcription factor (SPDEF) in Calu-3 airway epithelial cells. SPDEF is an integrator of goblet cell differentiation and pulmonary Th2 inflammation. ( C ) Low cycle numbers (20×) were chosen for quantification of expression by relating specific signals to expression of the housekeeper protein GAPDH. IL-13 (20 ng/mL) leads to upregulation of expression of MUC5AC, TMEM16A, and SPDEF. Niclosamide (Niclo, 1 µM, 72 h) strongly inhibits expression of MUC5AC, TMEM16A, and SPDEF. Mean ± SEM (number of assays). #,§ significant increase by IL-13 and inhibition by niclosamide, respectively ( p < 0.05, ANOVA).
Article Snippet: The primary antibody, anti-DOG1/TMEM16A (SP31, Novus Biologicals, USA) was incubated overnight at 4 °C.
Techniques: Expressing, Western Blot, Inhibition, Reverse Transcription Polymerase Chain Reaction, Cell Differentiation