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. 2014 Aug 18;9(8):e105185. doi: 10.1371/journal.pone.0105185

Figure 2. The deletion of the putative NLS of hARD1 blocks nuclear localization.

Figure 2

A. The structure and sequence comparison of the putative NLS in ARD1. The reported structure of yeast ARD1 is shown. The region corresponding to the NLS in hARD1 (a.a. 79–84, MRSYRH in yeast [21]) is colored yellow. B. Schematic presentation of an NLS deletion mutant of hARD1. Wild type hARD1 was tagged with GFP at its N-terminus (GFP-hARD1 WT), and the putative NLS was deleted (GFP-hARD1ΔN). C. Deletion of NLS compromised nuclear translocation of hARD1. HEK293T cells transfected with GFP-hARD1 WT and ΔN were subjected to nuclear/cytosolic fractionation and GFP-hARD1 levels were detected by anti-GFP antibody. Note the slight decrease in size of ΔN relative to WT. D. NLS-deleted hARD1 was unable to access the nucleus. GFP-hARD1 WT- and ΔN-expressing HeLa cells were visualized by fluorescence microscopy. Scale bar, 10 µm.