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. 2014 Dec 2;5(1):111–121. doi: 10.1534/g3.114.015800

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Copyright © 2015 Rogers and Rose

This is an open-access article distributed under the terms of the Creative Commons Attribution Unported License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Kar5p localizes to both nuclear envelope faces. Indicated CEN GFP1-10 plasmids were expressed in strain MY14667 (GFP11-mCherry-Pus1) or MY14668 (GFP11-mCherry-SCS2-TM). Green fluorescence indicates that GFP1-10 and GFP11 can colocalize and consequently fluoresce. Note that Sey1-GFP1-10 only fluoresces with the Scs2-TM marker and is excluded from the nucleus (arrowhead), demonstrating outer/inner nuclear envelope specificity in this assay. In Kar5-TM3-GFP1-10, note the bright fluorescence (arrowhead) near the shmoo tip when combined with GFP11-mCherry-Pus1. Because different fluorescent reporters were used (with differing expression levels), images are not displayed at the same brightness. Scale bars, 2 μm.