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. 2013 Oct 1;24(19):3133–3144. doi: 10.1091/mbc.E13-05-0269

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© 2013 Lipatova 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 2: — Excess ER accumulates in ypt1-1 mutant cells expressing fluorescently tagged ER proteins. (A) Excess ER accumulates in ypt1-1 mutant cells expressing Hmg1-mCherry. The ER protein Hmg1 was tagged on the chromosome with mCherry in wild-type and ypt1‑1 and ypt31∆/32ts mutant cells. Cells were visualized by live-cell fluorescence microscopy. Left to right, differential interference contrast (DIC), mCherry, and percentage of cells with aberrant Hmg1 structures (N, number of cells visualized). In wild-type and ypt31∆/32ts mutant cells, mCherry-tagged Hmg1 localizes to rings around nuclei, a typical ER staining pattern (Huh et al., 2003). On the other hand, excess ER structures accumulate in the majority of ypt1‑1 mutant cells. (B) Excess ER accumulates in ypt1-1 mutant cells expressing Sec61-mCherry. Sec61 was tagged on the chromosome with mCherry in wild-type and ypt1‑1 mutant cells. Cells were visualized by live-cell fluorescence microscopy. Left to right, DIC, mCherry, and percentage of cells with aberrant Sec61 structures (N, number of cells visualized). In wild-type cells, mCherry-tagged Sec61 localizes to rings around nuclei and underneath the PM, as previously seen for Sec61-GFP (Huh et al., 2003). In addition, excess ER structures accumulate in the majority of ypt1‑1 mutant cells. (C) Excess ER accumulates in ypt1-1 mutant cells expressing Nup60–yeast enhanced GFP (yEGFP) and Hmg1-mCherry. The nuclear pore protein Nup60 was tagged on the chromosome with yEGFP in wild type and ypt1‑1 mutant cells expressing Hmg1-mCherry. Cells were visualized by live-cell fluorescence microscopy. Left to right, DIC, GFP, mCherry, merge, percentage of cells with Nup60 other than the ring (N, number of cells visualized). In wild-type cells, Nup60-GFP localizes to rings around nuclei (Huh et al., 2003). About half of the ypt1-1 mutant cells (47%) contain structures other than the rings in which Nup60-GFP colocalizes with Hmg1-mCherry (92%). Arrows point to aberrant ER structures; bars, 5 μm. Results represent at least two independent experiments.