Figure 9.

Sites of classical endocytosis do not colocalize with MCC. (B and C) The plasma membrane distributions of Rvs161 (B) and Ede1 (C), markers of late and early endocytic steps, respectively, were tested for colocalization with the MCC marker Sur7. For comparison, localization of the MCC resident Nce102 was analyzed (A). Tangential confocal sections showing the cell surface are presented. Because of a high mobility of Rvs161 patches, a maximum intensity projection of 36 frames (5 s per frame) instead of a single frame is shown in B. In this arrangement, a higher number of Rvs161 patches could be localized toward the stable Sur7 pattern at the same time. The rate of colocalization was quantified by fluorescence intensity profiles (top diagrams and arrows in merge) and 2D scatter plots of the whole full resolution images (Fig. S4, available at http://www.jcb.org/cgi/content/full/jcb.200806035/DC1). For easy orientation in the scatter plots, real pixel colors were used. Note the diagonal orientation of the Nce102-derived scatter plot demonstrating the colocalization of red and green fluorescence signals and a clear separation of red and green pixels in the two other cases. Examples of Sur7 patches adjacent to endocytic sites are highlighted (arrowheads). Bar, 5 μm.