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. 2014 Apr 15;25(8):1202–1215. doi: 10.1091/mbc.E13-07-0430

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© 2014 Datta et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 9: — Nuclear localization of Ubc9 is resistant to the effects of H₂O₂ and progerin in CHO cells preadapted to oxidative stress. (A) Cells were treated with the indicated concentrations of H₂O₂ for 10 min, and endogenous Ubc9 and Ran were detected by IF. Scale bars, 20 μm. (B, C) Ubc9-Uba2 disulfide formation in HA-1 and OC14 cells. Cells treated with different concentrations of H₂O₂ for 10 min were analyzed by immunoblotting under nonreducing conditions. The electrophoretic position of the Ubc9-Uba2 disulfide (arrow) and a nonspecific cross-reaction (asterisk) are indicated. (D) Transfection of HA-lamin and HA-progerin into CHO cell lines, followed by detection of endogenous Ubc9 and Ran by IF. Lamin A and progerin expression was detected with anti-HA antibody. (E, F) Histograms (bin size, 0.5) showing Ran N/C and Ubc9 N/C in the HA-1(parental) and OC-14(resistant) cells transfected with progerin. Scale bars, 20 μm.