Figure 5: FALC activation during acute airway inflammation is IL-33 dependent.

(a) ELISA of IL-33 in the pleural lavage (pl Lav) of naive and Alt-treated C57BL/6 mice. Data are combined from two independent experiments, symbols represent individual mice, n=6 or 9 per group. (b) Representative whole-mount immunofluorescence staining of mediastinal FALCs from naive and Alt-treated BALB/c and Il1rl1^−/− mice showing IgM (green), Ki67 (red) and B220 (blue) and quantification of the percentage area of Ki67 staining within individual clusters, scale bar, 50 μm. (c,d) Flow cytometric analysis showing proliferating Ki67^high and Ki67^int (c, left panel) and quantification of % Ki67^high (c, right panel) and number (d) of B1a (first row), B1b (second row) and B2 (third row) cells from digested pericardial FALC cells and PLEC from naive and Alt-treated BALB/c and Il1rl1^−/− mice. (e) ELISA of total IgM in the pleural lavage (pl Lav), peritoneal lavage (p Lav) and serum of naive and Alt-treated BALB/c and Il1rl1^−/− mice. Data in c–e are combined from two independent experiments, symbols represent individual mice, n=8 or 10 per time point. (f) ELISA of total IgM in the supernatant of overnight culture of PLEC and digested FALC cells from naive and Alt-treated BALB/c mice. Data are combined from three independent experiments, symbols represent individual mice, n= 9 or 14 per time point. ns not significant, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 (normally distributed data analysed by one-way ANOVA with Sidak multiple comparisons post test, non-normally distributed data analysed using the Kruksal–Wallis test with Dunn’s multiple comparisons post test). Error bars represent mean with s.e.m. in all graphs.