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. 2013 Jun 15;24(12):2021–2033. doi: 10.1091/mbc.E13-03-0160

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© 2013 Castillon et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 5: — N-glycosylation is required for proper sorting of fully remodeled GPI-APs and lysoGPI-APs. (A) Orthogonal views of control cells expressing F-CD59 or F-CD59(N43A) grown on filters for 6 d. The signal intensity was measured by immunofluorescence staining of F-CD59 after fixation in 100% methanol at −20°C for 20 min and labeling with an anti-CD59 antibody. The means of apical sorting (percentage of apical signal/total signal) + SD for 20 cells from two independent experiments are shown. The micrographs are pseudocolored as in Figure 4B. (B) Polarized release of F-CD59 or F-CD59(N43A) into the apical (A) and basolateral (BL) chambers of PGAP3-HA cells grown on filters for 6 d during a 2-h chase after pulse labeling was analyzed using anti-FLAG M2 beads. The means of apical sorting (percentage of apical signal/total signal) + SD from five independent experiments are shown.