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. 2008 Aug;19(8):3404–3414. doi: 10.1091/mbc.E08-04-0354

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© 2008 by The American Society for Cell Biology

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Figure 3. — ER localization of malectin. (A) Schematic drawing of the FLAG-tagged malectin constructs used for transient transfection experiments. N-FLAG-malectin represents the full-length protein including a FLAG-tag (F) between the predicted N-terminal signal peptide (red, SP; AA 1-26) and the conserved lectin-like domain (blue, LLD; AA 27-213) that is followed by a hydrophobic C-terminal domain (yellow, HD; AA 255-274). The deletion construct ΔN-FLAG-malectin lacks the N-terminal SP. (B) Immunofluorescence analysis of U-2 OS cells for FLAG-tagged malectin (green) and the ER marker calnexin (red) showing that malectin and calnexin colocalize in ER structures (arrows indicate presence of both markers in the nuclear envelope). (C) Cytoplasmic localization of ΔN-FLAG-malectin after deletion of the predicted N-terminal signal peptide. Note that ΔN-FLAG-malectin also diffuses to the nucleus (asterisks) and can be found in protein aggregates (arrow heads). Bar, 10 μm.