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Fig. 3. Syndecan-4 is internalized with endogenous GPI-anchored proteins. (a) FcR-S4 expressing RFPES were labeled with FLAER, decorated by biotinylated niIgG and clustered by Cy-3-F(ab')2. (b) Five minutes after clustering, cells were placed on ice and the cell surface FcR-S4 was detected with streptavidin-Cy5. (c) FLAER-labeled GPI-anchored proteins. (d-f) Cell surface (d,f) and internalized (e,f) FcR-S4 demonstrate high level of colocalization with GPI-anchored proteins. Arrows in f point to examples of FcR-S4- and FLAER-labeled internalized vesicles. Scale bars, 10 µm.