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. 2008 Apr 25;283(17):11772–11784. doi: 10.1074/jbc.M707572200

FIGURE 7.

FIGURE 7.

Nef-eYFP and endogenous CI-MPR are mislocalized in PACS-2-/- MEFs. A, PACS-2+/+ and PACS-2-/- MEFs were transfected (Amaxa) with pNef-eYFP. After 18 h the cells were fixed and stained with anti-mannosidase II or anti-EEA1 (red) and visualized by confocal microscopy. Western blot (WB) in upper right-hand corner of PACS-2 in lysates from PACS-2+/+ and PACS-2-/- MEFs is shown. Inset, Nef-eYFP in dispersed EEA1-postive compartments in PACS-2-/- MEFs. B, PACS-2+/+ and PACS-2-/- MEFs were transfected (Amaxa) or not with pGalT-CFP. After 18 h the cells were fixed, stained with anti-EEA1 (red) or anti-CI-MPR 8738 (red in GalT-expressing cells; green in EEA1 co-stained cells (bottom)), and visualized by confocal microscopy. Inset, CI-MPR in EEA1-positive compartments. Morphometric analysis was performed as described under “Experimental Procedures.” Error bars represent mean ± S.D. from 20 cells per marker in four independent experiments for Nef-eYFP and three independent experiments for CI-MPR. Scale bar = 20 μm.