FIGURE 5.

Translocation of cPLA₂ and 5-LO in response to sPLA₂-X. Eosinophils were allowed to adhere to BSA-coated coverslips and treated with vehicle alone (Unstimulated), sPLA₂-X (100 nm), or fMLP (100 nm). The fMLP-stimulated cells served as a positive control. Cells were fixed, permeabilized, and immunostained with antibodies directed against cPLA₂ (A) and 5-LO (B). Fluorescence from the secondary Cy3-labeled antibody was visualized with confocal microscopy. A, in unstimulated cells, the cPLA₂ immunostaining was faint and diffuse, but the immunostaining increased following treatment of the cells with sPLA₂-X and localized in the cells in the perinuclear space as well as punctate cytoplasmic staining. Similar immunostaining was observed for eosinophils treated with fMLP as a positive control. B, the 5-LO immunostaining in unstimulated cells was also faint and diffuse, but the immunostaining increased following treatment of the cells with sPLA₂-X in both perinuclear and focal intracytoplasmic locations. Similar immunostaining for 5-LO was observed for eosinophils treated with fMLP as a positive control.