Fig. 2

Flagellin induces mucus secretion. Developing bronchospheres were treated with different PAMPs during the differentiation phase starting at day 2 after seeding. Bronchospheres were analyzed 30 days after seeding. a Immunofluorescence staining was performed for KRT5 (red channel) and MUC5B (green channel). Nuclei were stained with DAPI (blue). Scale bar: 100 μm. b Quantification of MUC5B staining. c Quantification of KRT5 staining. d Immunoblot analysis of lysates of bronchospheres cultured in the presence of flagellin, LPS, or control media probed with antibody to MUC5B or β-actin. e Immunohistochemistry was performed for MUC5AC. Scale bar: 20 μm. f Quantification of MUC5AC staining. Data were compared by unpaired Student’s t-test and are shown as the mean ± SEM from three independent experiments. *p < 0.05, **p < 0.01