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Fig. 5. Ionomycin-induced discharge of macropinosomes. Panel A shows the confocal merged images of DCs loaded for 10 minutes with Bds (green) before fixation, followed by EEA1 immunolabelling (red). The large co-localization of Bds and EEA1 (yellow) is similar to that in Fig. 1E. Panel B shows DCs as in A, but exposed for 1 minute to IONO before fixation and EEA1 immunolabelling. Notice the complete disappearance of the Bds-positive puncta with apparent no change in the puncta positive for EEA1 alone. Bar, 10 µm (A), also valid for B. Panel C shows mean ± s.d. of results on Bd-EEA1-positive puncta such as those in A and B, quantified morphometrically in at least ten fields. Notice that the Bd-EEA1-positive puncta, which before IONO were almost 45% of the whole EEA1-positive population, dropped to <3% in 1 minute of application of the ionophore. Panels D and E illustrate results such as those in A and B, respectively, but at the electron microscope level. The macropinosome vacuoles packed with Bds (arrows in D), similar to those of Fig. 1H,I, are no longer visible after 1 minute of IONO administration (E). Bar, 0.5 µm (D), also valid for E.
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