Fig. 2.
CD16 engagement induces the transient translocation of SHIP-1 to raft fraction.
Unstimulated or CD16-stimulated (2 minutes) cultured NK cells (3 × 108/sample) were fractionated by sucrose gradient centrifugation as described in “Materials and methods.” NK cells were stimulated either by anti-CD16 mAb plus secondary GAM (A-D, right) or by IgG- or IgG F(ab′)2–coupled polystyrene beads (E). Then, 8% (A-B) and 15% (C-D) SDS-PAGEs were loaded with samples from the same experiment. The 8% SDS-PAGE (E) was loaded with samples from a different experiment. The equivalent protein amount within the lanes was checked by Ponceau S red staining. Western blot analysis with anti-SHIP, antitubulin, anti-LAT or peroxidase-conjugated CTB was performed. Two experiments representative of each kind of stimulation are shown.