Fig. 2

Increased expression of SPDEF and decreased expression of FOXA2 in the bronchiolar epithelium of Adgrf5^−/− mice. a–d Immunofluorescence confocal microscopy was performed on lung sections from Adgrf5^−/− (a and c) and WT mice (b and d) at 30 weeks of age with an antibody specific for SPDEF (a and b, green in left panels, middle panels) or FOXA2 (c and d, green in left panels, middle panels). The nuclei were stained with Hoechst 33342 (blue in left panels, right panels). Scale bars, 20 μm. e FOXA2-negative bronchiolar epithelial cells and total bronchiolar epithelial cells were counted (total number of cells counted > 800). Results are expressed as percentage of FOXA2-negative cells relative to total bronchiolar epithelial cells, and are presented as the mean ± SEM (n = 6). ***p < 0.0001. f Whole lung lysates (20 μg of protein) from WT and Adgrf5^−/− mice (5, 12, and 30 weeks of age) were analyzed by western blotting using antibodies specific for SPDEF (top) and α-tubulin (bottom)