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. 2007 Feb 21;2(2):e237. doi: 10.1371/journal.pone.0000237

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Nielsen et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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MCF-7 cells stably expressing ectopic murine Podocalyxin were either transfected with VSV-tagged dominant negative (N-terminal) ezrin after formation of monolayers and appropriate localization of Podocalyxin and NHERF-1 (A, B) or transfected with VSV-tagged N'ezrin and then replated before immunostaining (C, D). (A–D) Immunolabeling of VSV-tagged N'ezrin (green), ectopic Podocalyxin (red), and endogenous NHERF-1 (blue). Purple represents apical colocalization of Podocalyxin and NHERF-1 in cells not expressing dominant negative ezrin (internal negative control); white represents apical colocalization of all three molecules in cells expressing dominant negative ezrin. Scale bars: 5 µm. (A, C) Projections of merged confocal stacks taken near the apical cell surface showing individual colors and merged images; (B, D) vertical slices of confocal stacks.