Fig. 7

Increased serum IgE levels and pulmonary mast cell accumulation in Adgrf5^−/− mice. a The levels of total serum IgE in WT and Adgrf5^−/−mice (3, 8, and 30 weeks of age) were determined by enzyme-linked immunosorbent assay (ELISA). Values are presented as the mean ± SEM (n = 3 for 3 weeks old, n = 5 for 8 weeks old, n = 4 for 30 weeks old). ***p < 0.001 vs. 30-week-old Adgrf5^−/− mice. b and c Alcian blue-periodic acid Schiff staining was performed on lung sections from WT (b) and Adgrf5^−/− mice (c) at 30 weeks of age. Nuclei (blue purple) were counter-stained with hematoxylin. Mucus and mast cells (arrows) were stained dark purple and light blue, respectively. Scale bars, 50 μm. d Whole lung lysates (30 μg of protein) from WT and Adgrf5^−/− mice (3, 5, 12, and 30 weeks of age) were analyzed by western blotting using antibodies specific for TPSAB1 (top) and α-tubulin (bottom). The band intensity was normalized to that of α-tubulin and is expressed as a value relative to that of corresponding WT mice. Values are presented as the mean ± SEM (n = 3). *p < 0.05 and ***p < 0.001. e–g Wright-Giemsa staining was performed on BAL cells from WT and Adgrf5^−/− mice (4 and 30 weeks of age). The numbers of macrophages (Mac), neutrophils (Neu), lymphocytes (Lym), and eosinophils (Eos) were counted. Results are expressed as the percentage of each inflammatory cell relative to total BAL cells, and are presented as the mean ± SEM (n = 3). *p < 0.05 and **p < 0.001. Representative images of BAL cells from 4-week-old WT and Adgrf5^−/− mice are shown in (f) and (g), respectively. The arrow indicates neutrophils. Scale bars, 50 μm. h Lung sections from Adgrf5^−/− mice at 10 weeks of age were stained with an Eosinophil-Mast Cell Stain Kit. Nuclei (blue purple) were counter-stained with hematoxylin. A representative image of an eosinophil (arrow) is shown. Scale bar, 50 μm