High ALDH expression on CD11cint CD11bhi LP eosinophils.

(A) Cells from LP were stained with anti-mouse CD11c, CD11b and Siglec-F Abs, and then CD11c+ CD11b¯ (P1), CD11c+ CD11b+ (P2), CD11c¯ CD11b+ side scatter (SSC)lo Siglec-F¯ (P3) and CD11c¯ CD11b+ SSChi Siglec-F+ cells (P4) were gated by flow cytometry.

(B) Cells from LP were stained with a panel of Abs and the indicated surface molecules of four populations were analyzed by flow cytometry. Filled histograms are the isotype control.

(C) Four subsets were sorted and ALDH activity of four subsets were determined by Aldefluor assay.

(D) Statistical summary of ALDH activity of four subsets. Data from experiments such as that shown in C. Bars show the means ± SD of three independent experiments.

(E) CD11b+ cells were isolated from the LP and stained with anti-mouse Siglec-F and CCR3 Abs, the percentage of Siglec-F+CCR3+ eosinophils were analyzed by flow cytometry.

(F) May-Grunwald-Giemsa staining of sorted CD11b+Siglec-F+CCR3+ cells from LP. Scale bar, 20 mm.