FIGURE 2.

DRM-Hs integrity is essential for FcγRIIa degradation and CIN85 recruitment. Neutrophils (20 × 10⁶ cells/ml) were preincubated in the presence or absence of 10 mm mβCD for 15 min at 37 °C before FcγRIIa cross-linking. A, whole cell lysates were probed by immunoblotting for FcγRIIa (CT10 antibody) and c-Cbl as a loading control. B, following mβCD preincubation, neutrophils were centrifuged and resuspended at 40 × 10⁶ cells/ml. Plasma membranes were prepared as described under “Experimental Procedures” and analyzed by immunoblotting for the presence of CIN85 and c-Cbl. The same blot was reprobed for flotillin-1 (loading control). The line graphs represent the means of densitometric ratios of three independent experiments. WB, Western blot. Error bars, S.E.