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. 2010 Dec 1;21(23):4227–4239. doi: 10.1091/mbc.E10-05-0449

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© 2010 by The American Society for Cell Biology

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 3. — Subcellular localization of SUMO-2 during arsenic treatment. (A) Characterization of HeLa YFP-SUMO-2 cell line. HeLa cells were stably transfected with a plasmid expressing a YFP-SUMO-2 fusion protein. Whole cells extracts from both parental HeLa and stable YFP-SUMO-2 cells were analyzed by SDS-PAGE followed by Western blot with rabbit anti-SUMO-2 and mouse anti-GFP antibodies. (B) HeLa YFP-SUMO-2 stable cell line was transfected with siRNA to RNF4 or a control nontargeting si RNA (NT) for 48 h before adding 1 μM of arsenic. Whole cell extracts were analyzed by SDS-PAGE followed by Western Blotting with rabbit anti-SUMO-2 and chicken anti-PML antibodies. Depletion of RNF4 was controlled with a rabbit anti-RNF4 antibody. (C) Subcellular localization of SUMO-2 during arsenic treatment. YFP-SUMO-2 HeLa cells were transfected with siRNA to RNF4 or a control nontargeting siRNA (NT) and YFP fluorescence measured for 18 h after the addition of 1 μM of arsenic. Images shows a projection of z-sections collected into YFP channel at selected time points. Bar, 5 μM.