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Fig. 7. MßCD treatment inhibits all CD82 signaling events. Jurkat cells, treated or not with 5 mM MßCD for 30 minutes, were stimulated for various times at 37°C on antibody-coated plates. 0, nonspecific mAbs; CD3s, suboptimal doses of OKT3 (0.5 µg ml–1); CD3o, optimal doses of OKT3 (20 µg ml–1); CD82, 50 µg ml–1 
C11. (A,B) 2x105 Jurkat cells were cultured for 1 hour on antibody-coated plates. (A) The number of adherent cells was evaluated by crystal violet staining, followed by densitometric analysis at 520 nm. (B) Adherent cells were fixed, permeabilized, stained with rhodamine-phalloidin and visualized by confocal microscopy. Bar, 10 µm. (C) 2x105 Jurkat cells were cultured for 10 minutes on antibody-coated plates and lysed in SDS sample buffer. Proteins were resolved by SDS-PAGE and tyrosine phosphorylations were evaluated by immunoblotting with the anti-phosphotyrosine antibody 4G10. Cell treatments and stimulations are indicated at the top. Arrows indicate proteins for which phosphorylation was reduced by MßCD treatment.
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