Figure 7.

Pulse–chase analysis of endosome to Weibel–Palade bodies transport. Cells were incubated in the presence of mAbs against CD63/lamp3 for 2 h (A) or P-selectin for 1 h (B) (pulse) and then further incubated in the absence of antibody for the indicated time period (chase). Anti-CD63/lamp3 antibodies were used at a concentration of 20 μg/ml in the medium and revealed by indirect immunofluorescence, whereas antibodies against P-selectin were directly conjugated with Alexa 568 and used at a concentration of 5 μg/ml. Cells were fixed and analyzed by double-immunofluorescence microscopy using antibodies against vWF. As in Figure 4, exposure times were adjusted so that labeled structures could be well resolved. Bar, 10 μm.