Fig. 3.

Treatment with PI3K p110δ inhibitor alters adhesion molecule expression and distribution by eosinophils. A: expression of adhesion molecules by vehicle- and IC87114-treated BM-Eos by flow cytometry using rat monoclonal antibodies (mAbs) against α4 (CD49), lymphocyte function associated antigen (LFA)-1 (CD11a), Mac-1 (CD11b), and CD62L. Depending on the mAb, rat IgG2a or 2b was used as the isotype-matched control. All antibodies were used at a final concentration of 5 μg/ml. B: immunofluorescence staining to evaluate distribution of Mac-1 and α4 on the cell surface of vehicle- or IC87114-treated eosinophils adhered to ICAM-1 and VCAM-1, respectively. Adhered cells were stained with goat anti-mouse α4 (5 μg/ml) or rat anti-mouse Mac-1 (10 μg/ml) followed by rhodamine-conjugated secondary antibodies. Goat IgG (for anti-α4) or rat IgG2b (for anti-Mac-1) was used as the isotype control. Magnification ×600. Data shown in A and B are representative of 3 independent experiments with eosinophils from 3 different mice.