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. 2019 Apr 25;8:e41331. doi: 10.7554/eLife.41331

Figure 7. Residues that play important roles in RanBP1 localization and nuclear export of cargo.

(A) mCherry tagged yRanBP1 was transfected alone or co-transfected with EGFP-yCRM1 or EGFP-yCRM1T753Q mutant, and visualized under microscope. (B) Quantification and statistical analysis of yRanBP1 localization in 7A. Ratio of nuclear yRanBP1 for each cell is calculated as its nuclear intensity divided by total cellular intensity. Error bars represent standard deviation of each set of data containing measurements from at least 30 cells. (C) mCherry tagged NESmut was transfected alone or co-transfected with EGFP-yCRM1 or EGFP-yCRM1T753Q mutant, and visualized under microscope. (D) Quantification and statistical analysis of NESmut localization in 7C. (E) Subcellular localization of endogenous RanBP1, NFκB and transfected mCherry-NLS under the treatment of siCRM1 and transfection of different EGFP-CRM1 constructs. mCherry-NLS is constructed as mCherry-NESPKI-MBP-NLSSV40 in pmCherry-C1 plasmid. (F–H) Quantification and statistical analysis of nuclear ratio of RanBP1, NFκB and mCherry-NLS in 7E.

Figure 7.

Figure 7—figure supplement 1. siRanBP1 does not change the subcellular localization of endogenous CRM1, NFκB and transfected GFP-NLS.

Figure 7—figure supplement 1.

(A) Subcellular localization of endogenous NFκB and the transfected GFP-NLS (constructed as GFP-NESPKI-MBP-NLSBPSV40) in the presence of siRanBP1 and transfection of different mCherry-RanBP1 constructs. (B–C) Quantification and statistical analysis of nuclear ratio of NFκB and GFP-NLS in A. (D) Subcellular localization of endogenous CRM1 and transfected GFP-NLS in the presence of siRanBP1 and transfection of different mCherry-RanBP1 constructs. (E) Western analysis shows that siCRM1 and siRanBP1 are effective.